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Backbone structure of a small helical integral membrane protein: A unique structural characterization

  1. Richard C. Page1,2,†,‡,
  2. Sangwon Lee3,‡,
  3. Jacob D. Moore1,2,
  4. Stanley J. Opella3,
  5. Timothy A. Cross1,2,*

Article first published online: 2 DEC 2008

DOI: 10.1002/pro.24

Issue

Protein Science

Protein Science

Volume 18, Issue 1, pages 134–146, January 2009

Additional Information

How to Cite

Page, R. C., Lee, S., Moore, J. D., Opella, S. J. and Cross, T. A. (2009), Backbone structure of a small helical integral membrane protein: A unique structural characterization. Protein Science, 18: 134–146. doi: 10.1002/pro.24

Author Information

  1. 1

    Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida 32306-4390

  2. 2

    National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida 32310

  3. 3

    Department of Chemistry and Biochemistry, University of California, San Diego, California 92093-0307

  1. †

    Department of Molecular Cardiology, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA

  1. ‡

    Richard C. Page and Sangwon Lee contributed equally to this manuscript.

*Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida 32306-4390

Publication History

  1. Issue published online: 16 DEC 2008
  2. Article first published online: 2 DEC 2008
  3. Accepted manuscript online: 2 DEC 2008 12:00AM EST
  4. Manuscript Accepted: 23 OCT 2008
  5. Manuscript Received: 25 JUL 2008

Funded by

  • National Institutes of Health. Grant Numbers: P01-GM64676, R01-AI23007, R01-AI073891, R01-GM075877, P41-EB002031
  • National High Magnetic Field Laboratory. Grant Number: DMR-0654118
  • American Heart Association predoctoral fellowship. Grant Number: 0615223B
  • National Institutes of Health predoctoral fellowship. Grant Number: F31NS054494

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Keywords:

  • integral membrane protein;
  • solution NMR;
  • detergent micelle;
  • residual dipolar couplings;
  • paramagnetic relaxation enhancement;
  • Mycobacterium tuberculosis

Abstract

The structural characterization of small integral membrane proteins pose a significant challenge for structural biology because of the multitude of molecular interactions between the protein and its heterogeneous environment. Here, the three-dimensional backbone structure of Rv1761c from Mycobacterium tuberculosis has been characterized using solution NMR spectroscopy and dodecylphosphocholine (DPC) micelles as a membrane mimetic environment. This 127 residue single transmembrane helix protein has a significant (10 kDa) C-terminal extramembranous domain. Five hundred and ninety distance, backbone dihedral, and orientational restraints were employed resulting in a 1.16 Å rmsd backbone structure with a transmembrane domain defined at 0.40 Å. The structure determination approach utilized residual dipolar coupling orientation data from partially aligned samples, long-range paramagnetic relaxation enhancement derived distances, and dihedral restraints from chemical shift indices to determine the global fold. This structural model of Rv1761c displays some influences by the membrane mimetic illustrating that the structure of these membrane proteins is dictated by a combination of the amino acid sequence and the protein's environment. These results demonstrate both the efficacy of the structural approach and the necessity to consider the biophysical properties of membrane mimetics when interpreting structural data of integral membrane proteins and, in particular, small integral membrane proteins.

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