Structure and function in bacteriorhodopsin: the role of the interhelical loops in the folding and stability of bacteriorhodopsin

Journal of Molecular Biology
J M KimH G Khorana

Abstract

Bacteriorhodopsin functions as a light-driven proton pump in Halobacterium salinarium. The functional protein consists of an apoprotein, bacterioopsin, with seven transmembrane alpha helices together with a covalently bound all-trans retinal chromophore. In order to study the role of the interhelical loop conformations in the structure and function of bacteriorhodopsin, we have constructed bacterioopsin genes where each loop is replaced, one at a time, by a peptide linker consisting of Gly-Gly-Ser- repeat sequences, which are believed to have flexible conformations. These mutant proteins have been expressed in Escherichia coli, purified and reconstituted with all-trans retinal in l-alpha-1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)/3-(3-cholamidopropyl)dimethylammonio-1-propane sulfonate (CHAPS)/SDS and l-alpha-1,2-dihexanoylphosphatidylcholine (DHPC)/DMPC/SDS micelles. Wild-type-like chromophore formation was observed in all the mutants containing single loop replacements. In the BC and FG mutants, an additional chromophore band with an absorption band at about 480 nm was observed, which was in equilibrium with the 550 nm, wild-type band. The position of the equilibrium depended on temperature, SDS and relative DMPC conc...Continue Reading

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Citations

Jan 15, 2009·Proceedings of the National Academy of Sciences of the United States of America·Paul Curnow, Paula J Booth
Oct 1, 2013·Molecular Biology and Evolution·Kristine A MackinDouglas L Theobald
May 5, 2004·Proceedings of the National Academy of Sciences of the United States of America·A J RaderJudith Klein-Seetharaman
Jun 5, 2014·Annual Review of Biophysics·Karen G Fleming
Jun 19, 2004·Molecular Membrane Biology·Paula J Booth, Stephen High
Apr 25, 2008·Protein Science : a Publication of the Protein Society·Damien Thévenin, Tzvetana Lazarova
Jun 3, 2009·The Journal of Physical Chemistry. B·Jing ZhangC Ted Lee
Mar 13, 2009·Biochemistry·Arianna RathCharles M Deber
Jan 24, 2006·Journal of Molecular Biology·Emma L R ComptonPaula J Booth
May 23, 2006·Archives of Biochemistry and Biophysics·Conceição A S A Minetti, David P Remeta
Jul 4, 2006·Current Opinion in Structural Biology·Paula J Booth, Paul Curnow
Mar 16, 2013·Biochemistry·Vincent G Nadeau, Charles M Deber
Jan 29, 2014·Journal of the American Chemical Society·Daryl B GoodVladimir Ladizhansky
Nov 1, 2007·Archives of Biochemistry and Biophysics·Ann Marie Stanley, Karen G Fleming
May 11, 2006·Chemical Reviews·Kevin R Mackenzie
Mar 18, 2009·Biophysical Journal·Oznur TastanHagai Meirovitch
Apr 7, 2010·Journal of Biotechnology·Oksana V NekrasovaMikhail P Kirpichnikov

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