Structure Determination of Membrane Proteins
EMSL Project ID
10301
Abstract
Despite the tremendous progress in structural biology in general, membrane proteins (MP’s) still represent an unconquered frontier. Over the years, my laboratory has become increasingly interested in this field and specifically the structure determination of MP’s by isotropic NMR solutions methods. Following my continuing, long-term collaboration with Dr. Chuck Sanders, now at Vanderbilt, on the structure of Diacylglycerol Kinase, we now participate in a program project headed by T. Cross, Florida State University focusing on the structure determination of MP’s from mycobacterium tuberculosis. A number of polytopic membrane proteins from TB have been successfully expressed, which are suitable for NMR structure determination and which can serve as vehicles for methodological advances in the structural characterization of MP’s, in addition to the genuine interest in their structure and function given the resurgence of TB.We seek two weeks of instrument time to begin experiments on Rv1616c, a 132 amino acid integral MP in sarcosyl micelles, and to continue our studies on Rv0011c, a 93 amino acid integral MP in DPC micelles. While the assignment of Rv0011c is in progress (backbone resonance assignment is > 50% complete) with data obtained at lower field and previously at the PNNL 800, we need additional data at elevated temperature to complete the resonance assignments. Even with the relatively small protein size, the added detergent weight in the protein detergent complexes pushes the spectroscopic behavior of Rv0011c and Rv1616c beyond the 30 kDa range, and the high magnetic field will particularly benefit these studies due the field dependence of TROSY techniques.
Project Details
Project type
Capability Research
Start Date
2004-10-15
End Date
2005-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members