Structure and function in rhodopsin: Mass spectrometric identification of the abnormal intradiscal disulfide bond in misfolded retinitis pigmentosa mutants

John Hwa, Judith Klein-Seetharaman, H. Gobind Khorana

Research output: Contribution to journalArticlepeer-review

62 Scopus citations

Abstract

Retinitis pigmentosa (RP) point mutations in both the intradiscal (ID) and transmembrane domains of rhodopsin cause partial or complete misfolding of rhodopsin, resulting in loss of 11-cis-retinal binding. Previous work has shown that misfolding is caused by the formation of a disulfide bond in the ID domain different from the native Cys-110-Cys-187 disulfide bond in native rhodopsin, Here we report on direct identification of the abnormal disulfide bond in misfolded RP mutants in the transmembrane domain by mass spectrometric analysis. This disulfide bond is between Cys-185 and Cys-187, the same as previously identified in misfolded RP mutations in the ID domain. The strategy described here should be generally applicable to identification of disulfide bonds in other integral membrane proteins.

Original languageEnglish (US)
Pages (from-to)4872-4876
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number9
DOIs
StatePublished - Apr 24 2001
Externally publishedYes

Keywords

  • Biotin-avidin affinity chromatography
  • G protein-coupled receptors
  • N-ethylmaleimide
  • Signal transduction
  • Transmembrane domain

ASJC Scopus subject areas

  • General

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