Proton movement and photointermediate kinetics in rhodopsin mutants.

Biochemistry
James W LewisDavid S Kliger

Abstract

The role of ionizable amino acid side chains in the bovine rhodopsin activation mechanism was studied in mutants E134Q, E134R/R135E, H211F, and E122Q. All mutants exhibited bathorhodopsin stability on the 30 ns to 1 micros time scale similar to that of the wild type. Lumirhodopsin decay was also similar to that of the wild type except for the H211F mutant where early decay (20 micros) to a second form of lumirhodopsin was seen, followed by formation of an extremely long-lived Meta I(480) product (34 ms), an intermediate which forms to a much reduced extent, if at all, in dodecyl maltoside suspensions of wild-type rhodopsin. A smaller amount of a similar long-lived Meta I(480) product was seen after photolysis of E122Q, but E134Q and E134R/R135Q displayed kinetics much more similar to those of the wild type under these conditions (i.e., no Meta I(480) product). These results support the idea that specific interaction of His211 and Glu122 plays a significant role in deprotonation of the retinylidene Schiff base and receptor activation. Proton uptake measurements using bromcresol purple showed that E122Q was qualitatively similar to wild-type rhodopsin, with at least one proton being released during lumirhodopsin decay per Meta I(...Continue Reading

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Citations

May 5, 2011·Biochemistry·Hisao TsukamotoDavid S Kliger
Mar 3, 2010·Annual Review of Biophysics·Steven O Smith
Jul 22, 2008·Journal of Molecular Biology·Supriyo BhattacharyaNagarajan Vaidehi
Nov 5, 2013·Biochimica Et Biophysica Acta·Andreyah PopeSteven O Smith
Jan 10, 2018·Frontiers in Molecular Biosciences·Kristina N WoodsJudith Klein-Seetharaman
Jan 9, 2018·Structural Dynamics·Krystel El HageMarkus Meuwly

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