Kinetics and pH dependence of light-induced deprotonation of the Schiff base of rhodopsin: possible coupling to proton uptake and formation of the active form of Meta II.
In this paper we first review what is known about the kinetics of Meta II formation, the role and stoichiometry of protons in Meta II formation, the kinetics of the light-induced changes of proton concentration, and the site of proton uptake. We then go on to compare the processes that lead to the deprotonation of the Schiff base in bacteriorhodopsin with rhodopsin. We point out that the similarity of the signs of the light-induced electrical signals from the two kinds of oriented pigment molecules could be explained by bacteriorhodopsin releasing a proton from its extracellular side while rhodopsin taking up a proton on its cytoplasmic side. We then examined the pH dependence of both the absorption spectrum of the unphotolyzed state and the amplitude and kinetics of Meta II formation in bovine rhodopsin. We also measured the effect of deuteration and azide on Meta II formation. We concluded that the pKa of the counter-ion to the Schiff base of bovine rhodopsin and of a surface residue that takes up a proton upon photolysis are both less than 4 in the unphotolyzed state. The data on pH dependence of Meta II formation indicated that the mechanisms involved are more complicated than just two sequential, isospectral forms of Meta II in the bleaching sequence. Finally we examined the evidence that, like in bacteriorhodopsin, the protonation of the Schiff bases's counter-ion (Glu113) is coupled to the changing of the pKa of a protonatable surface group, called Z for rhodopsin and tentatively assigned to Glu134. We conclude that there probably is such a coupling, leading to the formation of the active form of Meta II.
Kuwata
O
O
Center for Biophysics and Computational Biology, and Department of Biochemistry, University of Illinois, Urbana, Illinois 61801, USA.
Yuan
C
C
Misra
S
S
Govindjee
R
R
Ebrey
T G
TG
D001386:Azides / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D011522:Protons; D012243:Rhodopsin / Q000737:chemistry*; D012545:Schiff Bases
D001386:Azides; D011522:Protons; D012545:Schiff Bases; D012243:Rhodopsin