Dynamics of voltage profile in enzymatic ion transporters, demonstrated in electrokinetics of proton pumping rhodopsin.
H(+)-pumping rhodopsins mediate a primordial conversion of light to metabolic energy. Bacteriorhodopsin from Halobacterium salinarium is the first identified and (biochemically) best-studied H(+)-pumping rhodopsin. The electrical properties of H(+)-pumping rhodopsins, however, are known in more detail for the homolog Acetabularia rhodopsin, isolated from the eukaryotic green alga Acetabularia acetabulum. Based on data from Acetabularia rhodopsin we present a general reaction kinetic model of H(+)-pumping rhodopsins with only seven independent parameters, which fits the kinetic properties of photocurrents as functions of light, transmembrane voltage, internal and external pH, and time. The model describes fast photoisomerization of retinal with simultaneous H(+) transfer to an H(+) acceptor, reprotonation of retinal from the intracellular face via an H(+) donor, and proton release to the extracellular space via an H(+) release complex. The voltage sensitivities of the individual reaction steps and their temporal changes are treated here by a novel approach, whereby--as in an Ohmic voltage divider--the effective portions of the total transmembrane voltage decrease with the relative velocities of the individual reaction steps. This analysis quantitatively infers dynamic changes of the voltage profile and of the pK values of the H(+)-binding sites involved.
Hagedorn
Rolf
R
Humboldt-Universität zu Berlin, Fachbereich für Biologie, Experimentelle Biophysik, Berlin, Germany. rolf.hagedorn@rz.hu-berlin.de
Gradmann
Dietrich
D
Hegemann
Peter
P
D000076:Acetabularia / Q000201:enzymology / Q000378:metabolism*; D001436:Bacteriorhodopsins / Q000378:metabolism*; D004553:Electric Conductivity; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D008954:Models, Biological; D012680:Sensitivity and Specificity; D013997:Time Factors; D017186:Titrimetry
D001436:Bacteriorhodopsins