Electric-field-induced Schiff-base deprotonation in D85N mutant bacteriorhodopsin.
The application of an external electric field to dry films of Asp-85-->Asn mutant bacteriorhodopsin causes deprotonation of the Schiff base, resulting in a shift of the optical absorption maximum from 600 nm to 400 nm. This is in marked contrast to the case of wild-type bacteriorhodopsin films, in which electric fields produce a red-shifted product whose optical properties are similar to those of the acid-blue form of the protein. This difference is due to the much weaker binding of the Schiff-base proton in the mutant protein, as indicated by its low pK of approximately 9, as compared with the value pK approximately 13 in the wild type. Other bacteriorhodopsins with lowered Schiff-base pK values should also exhibit a field-induced shift in the protonation equilibrium of the Schiff base. We propose mechanisms to account for these observations.
Kolodner
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P
Bell Laboratories, Lucent Technologies Inc., Murray Hill, NJ 07974-0636, USA.
Lukashev
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EP
Ching
Y C
YC
Rousseau
D L
DL
D000595:Amino Acid Sequence; D001216:Asparagine; D001224:Aspartic Acid; D001436:Bacteriorhodopsins / Q000737:chemistry*; D004563:Electrochemistry / Q000379:methods; D014644:Genetic Variation; D006217:Halobacterium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D017354:Point Mutation; D012545:Schiff Bases; D013053:Spectrophotometry / Q000379:methods; D013997:Time Factors
D012545:Schiff Bases; D001224:Aspartic Acid; D001436:Bacteriorhodopsins; D001216:Asparagine