Difference: NaiderZerbe (6 vs. 7)

Revision 723 May 2008 - Main.OliverZerbe

META TOPICPARENT	name="ProposalEdit"

Set ALLOWTOPICVIEW = NmrfacGroup, NysbcadminGroup, FredNaider, OliverZerbe

META FORM	name="ProposalForm"
FORM FIELD Project Title	ProjectTitle	GPCR peptide structures
FORM FIELD Principle Investigator	PrincipleInvestigator	Fred Naider
FORM FIELD Grant Source	GrantSource
FORM FIELD Grant Number	GrantNumber
FORM FIELD Grant Dates	GrantDates
FORM FIELD Facilities Required	FacilitiesRequired	biochemical-facilities, solution-NMR
FORM FIELD Animal Origin	AnimalOrigin	neither
FORM FIELD Animal Species	AnimalSpecies
FORM FIELD Animal Protocol Num	AnimalProtocolNum
FORM FIELD Animal Protocol Date	AnimalProtocolDate
FORM FIELD Human Protocol Num	HumanProtocolNum
FORM FIELD Human Protocol Date	HumanProtocolDate

Changed:

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FORM FIELD Project Description	ProjectDescription

I would like to formally apply for measuring time on the 900 MHz instrument as discussed between Prof. Naider and Cowburn. The sample seems to be rather stable, and can certainly be stored in the fridge for periods of a couple of weeks. The 13C-NOESY needs to be measured at 320K, and for navigating through the 3D we would like to also have a constant-time 13C,1H HSQC. For the 2D 3-4 hours of measuring time are fine, for the 13C NOESY we expect to need approx. 3 days (16 scans) (at least). We would send the sample in the shigemi tube once the measuring slot has become clear. This will take approx. 3-4 days.

The sample is a doubly-labeled membrane protein (a GPCR fragment) in deuterated LPPG micelles. The protein concentration is approx. 400 uM, the detergent conc. is 200mM, pH about 6.

Pl. let me know if you need more information. I will provide more details on the sequences used here for the reference spectra, and the spectral widths, increments...etc

With kindest regards,

Oliver Zerbe

I think we would need the 2D ct-13C,1H HSQC to check for slight shift changes, and to also be guided through the 3D in the end. I believe the 13C-NOESY is all we need at the moment. We will adjust our assignments on that spectrum. The sample is in D2O at the moment, so we will miss the NOEs to amides, but will have much better quality in the aliphatic range. We will run 13C-HSQC here and could send it to the NY structural biology center electronically, so that the HSQC could be directly overlayed. We could check sample integrity here any time before. |

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|*FORM FIELD Project Description*|ProjectDescription|A 13C-resolved NOESY that was recorded recently for us at the NYSBC proved much superior to the 700 MHz data recorded here. In that light we would like to ask for a 13C NOESY centered at the aromatic carbons, and a corresponding constant-time HSQC (using 8 ms instead of 13.3) for navigation through the 3D. In addition to the better resolution in the proton dimension we also believe to benefit from fact that less problems occur from second order effects.|

FORM FIELD Preliminary Results	PreliminaryResults	Lower field spectra
FORM FIELD Staff Notes	StaffNotes	Prelim to 900 use for Fred Naider

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