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PMID: 3288985 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Aspartic acid substitutions affect proton translocation by bacteriorhodopsin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 12 ·1988-06-00 ·Pages 4148-52

Mogi T, Stern LJ, Marti T, Chao BH, Khorana HG

Abstract

We have substituted each of the aspartic acid residues in bacteriorhodopsin to determine their possible role in proton translocation by this protein. The aspartic acid residues were replaced by asparagines; in addition, Asp-85, -96, -115, and -112 were changed to glutamic acid and Asp-212 was also replaced by alanine. The mutant bacteriorhodopsin genes were expressed in Escherichia coli and the proteins were purified. The mutant proteins all regenerated bacteriorhodopsin-like chromophores when treated with a detergent-phospholipid mixture and retinal. However, the rates of regeneration of the chromophores and their lambda max varied widely. No support was obtained for the external point charge model for the opsin shift. The Asp-85----Asn mutant showed not detectable proton pumping, the Asp-96----Asn and Asp-212----Glu mutants showed less than 10% and the Asp-115----Glu mutant showed approximately equal to 30% of the normal proton pumping. The implications of these findings for possible mechanisms of proton translocation by bacteriorhodopsin are discussed.

MeSH Terms
Amino Acid Sequence Aspartic Acid Bacteriorhodopsins/genetics,metabolism Escherichia coli/genetics Halobacterium/genetics,metabolism Molecular Sequence Data Mutation Plasmids Protein Conformation
Chemicals
Aspartic Acid Bacteriorhodopsins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mogi T
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Stern L J
Marti T
Chao B H
Khorana H G
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27 references, click to expand
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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.85.12.4148
Published
1988-06-00
Pages
4148-52
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC280383
Subset
IM
Grants
NIGMS NIH HHS · R01GM28289-07 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.12.4148 PMC Full Text View journal details
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