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PMID: 3186735 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cysteine residues 110 and 187 are essential for the formation of correct structure in bovine rhodopsin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 22 ·1988-11-00 ·Pages 8459-63

Karnik SS, Sakmar TP, Chen HB, Khorana HG

Abstract

To investigate the role of different cysteine residues in bovine rhodopsin, a series of mutants were prepared in which the cysteine residues were systematically replaced by serines. The mutant genes were expressed in monkey kidney cells (COS-1) and the mutant opsins were evaluated for their levels of expression, glycosylation patterns, and ability to form the chromophore characteristic of rhodopsin and to activate transducin. Substitution of the three cytoplasmic cysteines (Cys-316, Cys-322, and Cys-323) and the four membrane-embedded cysteines (Cys-140, Cys-167, Cys-222, and Cys-264) produced proteins with wild-type phenotype. Also, single substitutions of Cys-185 gave rise to a wild-type phenotype. In contrast, substitution of the three intradiscal cysteines (Cys-110, Cys-185, and Cys-187) or single substitution of Cys-110 or Cys-187 gave proteins that were expressed at reduced levels, glycosylated abnormally, and unable to bind 11-cis-retinal. Thus, of the 10 cysteines in bovine rhodopsin, only intradiscal Cys-110 and Cys-187 are essential for the correct tertiary structure of the protein.

MeSH Terms
Amino Acid Sequence Animals Cattle Cysteine Molecular Sequence Data Mutation Phenotype Protein Conformation Retina/metabolism Retinal Pigments/genetics Rhodopsin/genetics,isolation & purification Spectrophotometry Transducin/isolation & purification
Chemicals
Retinal Pigments Rhodopsin Transducin Cysteine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Karnik S S
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Sakmar T P
Chen H B
Khorana H G
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.22.8459
Published
1988-11-00
Pages
8459-63
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC282477
Subset
IM
Grants
NIGMS NIH HHS · 5 F32 GM11305 · United States
NIGMS NIH HHS · GM28289-08 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.22.8459 PMC Full Text View journal details
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