Difference: JamesKempf (1 vs. 22)

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

TIME	PLACE	ACTIVITY
10:30A -	CCNY Marshak 1208B	RuthStark , RS will arrange for JK to go next
11:30A -	CCNY 325 C	CarlosMeriles, CM will arrange for JK to go next
12:30 -	A-11	setup for talk, lunch with affiliates/ staff
1:30P - 2:40P	A-11	Talk and discussion
2:40P - 3:30 P	A-20	ArthurPalmer
3:30P - 4:00 P	A-20	ShibaniBhattacharya
4:00P -	Sezz Medi	light dinner, contact DC if interested
5:00P -	to LGA	DC will drive JK to LGA
8:30A	Foyer NYSBC	DavidCowburn and / or MichaelGoger greet JK
8:30A - 9:30A	A-10 and tour	tour of NYSBC, breakfast / coffee etc
9:30A -	A-20 NYSBC	NMR staff and BorisItin, BI will walk JK to CCNY and Stark

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Schedule for talk on Oct 21 2009, 1:30 PM NYSBC

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

Abstract for talk on Oct 21, 2009

• Dynamic, Structural and Electrostatic Insights from NMR Spectroscopy
• Jim Kempf
• Department of Chemistry & Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180

Nuclear Magnetic Resonance (NMR) spectroscopy is well known as powerful probe of biomolecular structure and intramolecular motions. Unique mechanistic insights are available through complementary NMR studies of dynamics and biochemical assays of kinetics. My group has used this approach to explore the influence of glycosylation on the function of RNase A. Glycosylation (i.e., oligosaccharide attachment) is nature’s most frequent modification of proteins. About 50% of human proteins are glycosylated, and yet only a small handful of known protein structures include an oligosaccharide. Furthermore, functional regulation by glycosylation is known but poorly understood. We developed a simple approach to circumvent the challenges preventing widespread NMR studies of glycoproteins. With this, we are exploring the mechanism of glycosylation-induced functional loss in RNase A. Our results show a subtle effect where glycosylation alters intramolecular dynamics that are known to be critical to function of the unmodified enzyme. These are essential steps forward both for application of NMR to glycoproteins and for understanding modes of glycosylation-modulated function.

In a distinct project, we aim to add new capabilities to NMR as a probe of the electrostatic environment within biomolecules. Few methods are available to characterize the intramolecular E fields that govern diverse biochemical processes from photosynthesis to membrane channel transport to enzyme catalysis. Spectral parameters of NMR are unquestionably responsive to electrostatic influence, particularly in the face of the ~10 MV/cm fields common within biomolecules. Unfortunately, applied E fields are impractical beyond ~0.1-1 MV/cm. With that limitation, we predict NMR Stark effects are hopelessly small relative to spectral resolution in the solid state. Thus traditional approaches cannot quantify NMR Stark effects or meet our goal of developing calibrated NMR probes of intramolecular E fields. To overcome this challenge, we provide a POWER (perturbations observed with enhanced resolution) NMR technique that can resolve NMR Stark effects for CO groups at applied E fields of < 0.1 MV/cm. The POWER approach provides needed resolution by removing all spectral contributions other than those from an applied perturbation, e.g., an E field. I will present the first-ever experimental observation of NMR Stark effects in this context, as well as empirical and theoretical progress towards application to carbonyls on the protein backbone and small-molecule protein ligands.

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

My Links

• NysbcIntro - your introduction to NYSBC
• NetLiterate - short introduction to the style of this intranet

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

My Links

• NysbcIntro - your introduction to NYSBC
• NetLiterate - short introduction to the style of this intranet

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

My Links

• NysbcIntro - your introduction to NYSBC
• NetLiterate - short introduction to the style of this intranet

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools

My Links

• NysbcIntro - your introduction to NYSBC
• NetLiterate - short introduction to the style of this intranet

Personal Preferences

Uncomment preferences variables to activate them (remove the #-sign). Help and details on preferences variables are available in TWikiPreferences.

• Show tool-tip topic info on mouse-over of WikiWord links, on or off:
  • #Set LINKTOOLTIPINFO = off
• Horizontal size of text edit box:
  • #Set EDITBOXWIDTH = 70
• Vertical size of text edit box:
  • #Set EDITBOXHEIGHT = 22
• Style of text edit box. width: 99% for full window width (default), width: auto to disable.
  • #Set EDITBOXSTYLE = width: 99%
• Write protect your home page: (set it to your WikiName)
  • #Set ALLOWTOPICCHANGE = JamesKempf

Related Topics

• ChangePassword for changing your password
• ChangeEmailAddress for changing your email address
• TWikiUsers has a list of other Intranet users
• UserDocumentationCategory is a list of Intranet user documentation
• UserToolsCategory lists all Intranet user tools