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Multidimensional High-Field Magic-Angle Spinning NMR of Membrane and Nanocrystalline Proteins


EMSL Project ID
26716

Abstract

We aim to extend ongoing studies of membrane protein structure at the University of Illinois, conducted on 500 and 750 MHz instruments in 3.2 mm magic-angle spinning rotors, to the 900 MHz BioSolids instrumental capabilities at EMSL. Both nanocrystalline model proteins (GB1, DsbA) and membrane proteins (DsbB, cytochrome bo3 oxidase) are available. High-yield bacterial expression systems for isotopically labeled proteins have been developed at Illinois and initial data acquired. We expect that the resolution and sensitivity at 900 MHz will benefit from the quadratic dependence of sensitivity and resolution of BioSolids experiments demonstrated by preliminary data. This will permit chemical shift assignments to be performed in highly congested regions of the DsbB and cytochrome oxidase spectra. These efforts will thereby provide unique types of data sets not possible at Illinois, which will complement and extend ongoing scientific efforts in the important area of membrane protein structure determination.

Project Details

Project type
Capability Research
Start Date
2007-10-01
End Date
2008-10-02
Status
Closed

Team

Principal Investigator

Chad Rienstra
Institution
University of Illinois at Urbana-Champaign

Team Members

Lindsay Sperling
Institution
University of Illinois at Urbana-Champaign

Andrew Nieuwkoop
Institution
University of Illinois at Urbana-Champaign

Related Publications

Sperling LJ, AJ Nieuwkoop, AS Lipton, DA Berthold, and CM Rienstra. 2010. "High Resolution NMR Spectroscopy of Nanocrystalline Proteins at Ultra-High Magnetic Field." Journal of Biomolecular NMR 46(2):149-155. doi:10.1007/s10858-009-9389-9