Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS_Quarterly_Report-7-1-2011.docx" size="50566" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 1/1/2011" date="1309466991" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" user="Main.DavidStokes" version=""

Added:

>
>

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 1/1/2012" date="1340643752" name="TEMIMPS-quarterly-report-1-2012.doc" path="TEMIMPS-quarterly-report-1-2012.doc" size="33792" user="Main.DavidStokes" version="1"

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Added:

>
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META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 10/1/2011" date="1340643715" name="TEMIMPSQuarterlyReport-Oct-2011.doc" path="TEMIMPSQuarterlyReport-Oct-2011.doc" size="45568" user="Main.DavidStokes" version="1"
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2012" date="1340643778" name="TEMIMPS-quarterly-report-4-2012.doc" path="TEMIMPS-quarterly-report-4-2012.doc" size="59904" user="Main.DavidStokes" version="1"
META FILEATTACHMENT	attr="" autoattached="1" comment="Annual Report 5/1/2012" date="1340643822" name="TEMIMPS_progress_report_2012.pdf" path="TEMIMPS_progress_report_2012.pdf" size="1492133" user="Main.DavidStokes" version="1"

Revision 1230 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Quarterly Report 7/1/2011

Changed:

<
<

Annual Report: May 1, 2011

>
>

Annual Report: May 1, 2011

Deleted:

<
<

* TEMIMPS-progress-report-2011.pdf: progress report 2011

Quarterly Report 4/1/2011

Quarterly Report 1/1/2011

d79 1

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS_Quarterly_Report-7-1-2011.docx" size="50566" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 1/1/2011" date="1309466991" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 1130 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Changed:

<
<

>
>

Quarterly Report 7/1/2011

Deleted:

<
<

TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

Annual Report: May 1, 2011

* TEMIMPS-progress-report-2011.pdf: progress report 2011

Changed:

<
<

TEMIMPSQuarterlyReport-4-2011.docx: Quarterly Report 4/1/2011

>
>

Quarterly Report 4/1/2011

Quarterly Report 1/1/2011

Deleted:

<
<

Quarterly-Report-1-2011.docx: Quarterly Report 1/1/2011

d79 1

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS_Quarterly_Report-7-1-2011.docx" size="50566" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 1/1/2011" date="1309466991" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 1030 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Added:

>
>

TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

Annual Report: May 1, 2011

* TEMIMPS-progress-report-2011.pdf: progress report 2011

Added:

>
>

TEMIMPSQuarterlyReport-4-2011.docx: Quarterly Report 4/1/2011

Quarterly-Report-1-2011.docx: Quarterly Report 1/1/2011

d79 1

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

Deleted:

<
<

* TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

Deleted:

<
<

* Quarterly-Report-1-2011.docx: Quarterly Report 1/1/2011

Deleted:

<
<

* TEMIMPSQuarterlyReport-4-2011.docx: Quarterly Report 4/1/2011

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS_Quarterly_Report-7-1-2011.docx" size="50566" user="Main.DavidStokes" version=""

Added:

>
>

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 1/1/2011" date="1309466991" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" user="Main.DavidStokes" version=""

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""

Deleted:

<
<

META FILEATTACHMENT	attachment="TEMIMPSQuarterlyReport-4-2011.docx" attr="" comment="Quarterly Report 4/1/2011" date="1309467024" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" stream="TEMIMPSQuarterlyReport-4-2011.docx" user="Main.DavidStokes" version="0"

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 930 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Annual Report: May 1, 2011

* TEMIMPS-progress-report-2011.pdf: progress report 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

* TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

* Quarterly-Report-1-2011.docx: Quarterly Report 1/1/2011

Added:

>
>

* TEMIMPSQuarterlyReport-4-2011.docx: Quarterly Report 4/1/2011

Added:

>
>

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS_Quarterly_Report-7-1-2011.docx" size="50566" user="Main.DavidStokes" version=""

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""

Changed:

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META FILEATTACHMENT	attachment="Quarterly-Report-1-2011.docx" attr="" comment="Quarterly Report 1/1/2011" date="1309466990" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" stream="Quarterly-Report-1-2011.docx" user="Main.DavidStokes" version="0"

>
>

META FILEATTACHMENT	attachment="TEMIMPSQuarterlyReport-4-2011.docx" attr="" comment="Quarterly Report 4/1/2011" date="1309467024" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" stream="TEMIMPSQuarterlyReport-4-2011.docx" user="Main.DavidStokes" version="0"

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 830 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Annual Report: May 1, 2011

* TEMIMPS-progress-report-2011.pdf: progress report 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

* TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

Changed:

<
<

META FILEATTACHMENT	attachment="TEMIMPS-progress-report-2011.pdf" attr="" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" stream="TEMIMPS-progress-report-2011.pdf" user="Main.DavidStokes" version="0"
META FILEATTACHMENT	attachment="TEMIMPS_Quarterly_Report-7-1-2011.docx" attr="" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS Quarterly Report-7-1-2011.docx" size="50566" stream="TEMIMPS Quarterly Report-7-1-2011.docx" user="Main.DavidStokes" version="0"

>
>

* Quarterly-Report-1-2011.docx: Quarterly Report 1/1/2011

Added:

>
>

META FILEATTACHMENT	attr="" autoattached="1" comment="Quarterly Report 4/1/2011" date="1309467025" name="TEMIMPSQuarterlyReport-4-2011.docx" path="TEMIMPSQuarterlyReport-4-2011.docx" size="18457" user="Main.DavidStokes" version=""
META FILEATTACHMENT	attachment="Quarterly-Report-1-2011.docx" attr="" comment="Quarterly Report 1/1/2011" date="1309466990" name="Quarterly-Report-1-2011.docx" path="Quarterly-Report-1-2011.docx" size="13471" stream="Quarterly-Report-1-2011.docx" user="Main.DavidStokes" version="0"

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 730 Jun 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Annual Report: May 1, 2011

* TEMIMPS-progress-report-2011.pdf: progress report 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

Added:

>
>

* TEMIMPS_Quarterly_Report-7-1-2011.docx: Quarterly Report 7/1/2011

META FILEATTACHMENT	attachment="TEMIMPS-progress-report-2011.pdf" attr="" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" stream="TEMIMPS-progress-report-2011.pdf" user="Main.DavidStokes" version="0"

Added:

>
>

META FILEATTACHMENT	attachment="TEMIMPS_Quarterly_Report-7-1-2011.docx" attr="" comment="Quarterly Report 7/1/2011" date="1309466938" name="TEMIMPS_Quarterly_Report-7-1-2011.docx" path="TEMIMPS Quarterly Report-7-1-2011.docx" size="50566" stream="TEMIMPS Quarterly Report-7-1-2011.docx" user="Main.DavidStokes" version="0"

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 623 May 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Annual Report: May 1, 2011

Added:

>
>

* TEMIMPS-progress-report-2011.pdf: progress report 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

Deleted:

<
<

* TEMIMPS-progress-report-2011.pdf: progress report 2011

META FILEATTACHMENT	attachment="TEMIMPS-progress-report-2011.pdf" attr="" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" stream="TEMIMPS-progress-report-2011.pdf" user="Main.DavidStokes" version="0"
META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 523 May 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Annual Report: May 1, 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

Added:

>
>

* TEMIMPS-progress-report-2011.pdf: progress report 2011

META FILEATTACHMENT	attachment="TEMIMPS-progress-report-2011.pdf" attr="" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" stream="TEMIMPS-progress-report-2011.pdf" user="Main.DavidStokes" version="0"

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 423 May 2011 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Added:

>
>

Annual Report: May 1, 2011

PSI Annual Meeting: Dec 8-9, 2010

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 318 Nov 2010 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Deleted:

<
<

PSI Annual Meeting: Dec 8-9, 2010

Changed:

<
<

Goals for 2011

>
>

TEMIMPS Goals for 2011

Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets
- Establish pipeline for protein production from bacteria, yeast and insect cells
- Establish standard conditions for screening 2D crystallization of novel targets
- Implement methods for detergent removal/crystallization
  - microfluidic 96-well dialysis block
  - robotic micropipettor for incremental addition of cyclodextrins
- Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
- Demonstrate utility of phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods

Targets
- P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)

Changed:

<
<

Goals for next 5 years

>
>

TEMIMPS Goals for next 5 years

Targets
- Existing crystals
  - AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  - AQP0: Interaction of channel with native lipids
  - KvAP^? channel: Structure of membrane bound channel to address gating
- Novel targets
  - GPCRs: cannabinoid, seratonin
  - TRP channnels: TRPA1 pain receptor
  - Rhomboids
  - secondary transporters: Heme transporter FLVCR
- Oligomeric proteins
  - gamma-secretase
  - Kdp (ATP-dependent P-type potassium pump)
- Complexes
  - G-protein in complex with GIRK channel
  - AQP2 in complex with AKAP79 adaptor
  - Two-component system (PhoP^?/PhoQ)

Technologies
- establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers
  - automation of crystallization
  - automation of imaging
  - integration with database
  - general enough and affordable to interest other EM groups
- develop software for automated data collection
  - low-dose imaging
  - electron diffraction
- improve software for structure determination
  - novel methods for image analysis of poorly ordered crystals
  - more robust/automated analysis of electron diffraction

Protocols
- define important parameters affecting 2D crystallization
  - define matrix of conditions for crystallization screen
- extend methods for structure determination by electron crystallography
  - molecular replacement
  - phase extension
  - density modification

Notable achievements

none to date

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 218 Nov 2010 - Main.DavidStokes

   META TOPICPARENT   name="TemimpsGroup"
-  META TOPICPARENT
+ name="TemimpsGroup"
-<
<
->
>
+ PSI Annual Meeting: Dec 8-9, 2010
-<
<
+ Goals
  Goals for 2011
-<
<
+ Targets 
 Aqp2: expression, purification, crystallization, data collection for constructs mimicking phosphorylated, dephosphorylated state
  P2A3 Zn transporter: 3D reconstruction and comparison of membrane bound form with X-ray structure of detergent-solubilzed homolog (YiiP^?)
  Crystallization screening: 
 
 
 
  Technologies 
 Develop automated sample screening technologies
  Implement phase extension methods for electron crystallography
->
>
+ Technology development will dominate 2011 as we establish the tools necessary for attacking our protein targets 
 Establish pipeline for protein production from bacteria, yeast and insect cells
  Establish standard conditions for screening 2D crystallization of novel targets
  Implement methods for detergent removal/crystallization  
 microfluidic 96-well dialysis block
  robotic micropipettor for incremental addition of cyclodextrins 
 
  Develop software for automated imaging and evaluation of crystallization trials by electron microscopy
->
>
+ Demonstrate utility of phase extension methods for electron crystallography
  Implement iterative helical reconstruction methods
-<
<
+ Develop microfluidic 96-well dialysis chamber
->
>
+ Targets 
 P2A3 Zn transporter: Comparison of membrane bound form with X-ray structure of detergent-solubilized homolog (YiiP^?)
  Goals for next 5 years
->
>
  Targets
-<
<
+ Structure of complex
->
>
+ Existing crystals
->
>
+ AQP2: Comparison of constructs mimicking phosphorylated/dephosphorylated state
  AQP0: Interaction of channel with native lipids
  KvAP^? channel: Structure of membrane bound channel to address gating
 
  Novel targets 
 GPCRs: cannabinoid, seratonin
  TRP channnels: TRPA1 pain receptor
  Rhomboids
  secondary transporters: Heme transporter FLVCR
 
  Oligomeric proteins 
 gamma-secretase
  Kdp (ATP-dependent P-type potassium pump)
 
  Complexes
  G-protein in complex with GIRK channel
-<
<
+ tetraspanin adhesion protein complex
->
>
+ AQP2 in complex with AKAP79 adaptor
  Two-component system (PhoP^?/PhoQ)
-<
<
+ Structure of multi-subunit enzyme 
 gamma-secretase
  Kdp (ATP-dependent pump)
 
  Novel structures 
 GPCRs
  TRP channnels
  Rhomboids
  Heme transporter
  Technologies
-<
<
+ improved software for structure determination
  high-throughput pipeline for crystallization screening
  define physical parameters influencing crystallization
->
>
+ establish general high-throughput pipeline for screening 2D crystallization within lipid bilayers 
 automation of crystallization
  automation of imaging
->
>
+ integration with database
  general enough and affordable to interest other EM groups
 
  develop software for automated data collection 
 low-dose imaging
  electron diffraction
 
  improve software for structure determination 
 novel methods for image analysis of poorly ordered crystals
  more robust/automated analysis of electron diffraction
 
 
 
 

 Protocols 
 define important parameters affecting 2D crystallization
  define matrix of conditions for crystallization screen
->
>
+ extend methods for structure determination by electron crystallography 
 molecular replacement 
  phase extension
  density modification
  Notable achievements
->
>
+ none to date
-<
<
  Set ALLOWTOPICVIEW = 
 

-- DavidStokes - 14 Nov 2010

  META FILEATTACHMENT   attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""
-  META FILEATTACHMENT
+ attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

Revision 114 Nov 2010 - Main.DavidStokes

META TOPICPARENT	name="TemimpsGroup"

Goals

Goals for 2011

Targets
- Aqp2: expression, purification, crystallization, data collection for constructs mimicking phosphorylated, dephosphorylated state
- P2A3 Zn transporter: 3D reconstruction and comparison of membrane bound form with X-ray structure of detergent-solubilzed homolog (YiiP^?)
- Crystallization screening:
Technologies
- Develop automated sample screening technologies
- Implement phase extension methods for electron crystallography
- Implement iterative helical reconstruction methods
- Develop microfluidic 96-well dialysis chamber

Goals for next 5 years

Targets
- Structure of complex
  - G-protein in complex with GIRK channel
  - tetraspanin adhesion protein complex
  - Two-component system (PhoP^?/PhoQ)
- Structure of multi-subunit enzyme
  - gamma-secretase
  - Kdp (ATP-dependent pump)
- Novel structures
  - GPCRs
  - TRP channnels
  - Rhomboids
  - Heme transporter

Technologies
- improved software for structure determination
- high-throughput pipeline for crystallization screening
- define physical parameters influencing crystallization
  - define matrix of conditions for crystallization screen

Notable achievements

Set ALLOWTOPICVIEW =

-- DavidStokes - 14 Nov 2010

META FILEATTACHMENT	attr="" autoattached="1" comment="progress report 2011" date="1306168508" name="TEMIMPS-progress-report-2011.pdf" path="TEMIMPS-progress-report-2011.pdf" size="3762182" user="Main.DavidStokes" version=""

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