The objective of the proposed research is to demonstrate methods to assign chemical shifts and shift anisotrophies on uniformly labeled membrane proteins in the solid state. This is a challenging project however, no uniformly 13C, 15N labeled protein of even moderate size has yet been fully assigned in the solid state. There have been significant improvements in NMR methodology (pulse sequence, hardware) which point to this as a possibility. In particular, dipolar recoupling techniques with higher efficiencies, advanced decoupling methods, probes capable of 25-30 kHz MAS and 200 kHz decoupling fields, and widebore magnets with 1H frequencies of 750 MHz should lead to full sold-state NMR structure determinations of membrane proteins. The chemical shifts in the retinal binding pocket will be particularly interesting to analyze, given the significant interactions between the sidechains and retinal. These shifts will be used along with quantum chemical methods to analyze in detail the mechanism by which the proton is pumped uniaxially.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM020818-02
Application #
6518893
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Whitmarsh, John
Project Start
2001-04-01
Project End
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
2
Fiscal Year
2002
Total Cost
$44,212
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Internal Medicine/Medicine
Type
Schools of Arts and Sciences
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Su, Wei; Slevin, Michael K; Marzluff, William F et al. (2016) Synthetic mRNA with Superior Properties that Mimics the Intracellular Fates of Natural Histone mRNA. Methods Mol Biol 1428:93-114