Identification of core amino acids stabilizing rhodopsin

A. J. Rader, Gülsüm Anderson, Basak Isin, H. Gobind Khorana, Ivet Bahar, Judith Klein-Seetharaman

Research output: Contribution to journalArticlepeer-review

124 Scopus citations

Abstract

Rhodopsin is the only G protein-coupled receptor (GPCR) whose 3D structure is known; therefore, it serves as a prototype for studies of the GPCR family of proteins. Rhodopsin dysfunction has been linked to misfolding, caused by chemical modifications that affect the naturally occurring disulfide bond between C110 and C187. Here, we identify the structural elements that stabilize rhodopsin by computational analysis of the rhodopsin structure and comparison with data from previous in vitro mutational studies. We simulate the thermal unfolding of rhodopsin by breaking the native-state hydrogen bonds sequentially in the order of their relative strength, using the recently developed Floppy Inclusion and Rigid Substructure Topography (FIRST) method [Jacobs, D. J., Rader,A. J., Kuhn, L. A. & Thorpe, M. F. (2001) Proteins 44, 150-165]. Residues most stable under thermal denaturation are part of a core, which is assumed to be important for the formation and stability of folded rhodopsin. This core includes the C110-C187 disulfide bond at the center of residues forming the interface between the transmembrane and the extracellular domains near the retinal binding pocket. Fast mode analysis of rhodopsin using the Gaussian network model also identifies the disulfide bond and the retinal ligand binding pocket to be the most rigid region in rhodopsin. Experiments confirm that 90% of the amino acids predicted by the FIRST method to be part of the core cause misfolding upon mutation. The observed high degree of conservation (78.9%) of this disulfide bond across all GPCR classes suggests that it is critical for the stability and function of GPCRs.

Original languageEnglish (US)
Pages (from-to)7246-7251
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number19
DOIs
StatePublished - May 11 2004
Externally publishedYes

Keywords

  • Folding
  • G protein-coupled receptor
  • Membrane protein
  • Network models
  • Simulation

ASJC Scopus subject areas

  • General

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