Proton transfer reactions in native and deionized bacteriorhodopsin upon delipidation and monomerization.
We have investigated the role of the native lipids on bacteriorhodopsin (bR) proton transfer and their connection with the cation-binding role. We observe that both the efficiency of M formation and the kinetics of M rise and decay depend on the lipids and lattice but, as the lipids are removed, the cation binding is a much less important factor for the proton pumping function. Upon 75% delipidation using 3-[(cholamidopropyl)dimethylammonio]-propanesulfonate (CHAPS), the M formation and decay kinetics are much slower than the native, and the efficiency of M formation is approximately 30%-40% that of the native. Upon monomerization of bR by Trition X-100, the efficiency of M recovers close to that of the native (depending on pH), M formation is approximately 10 times faster, and M decay kinetics are comparable to native at pH 7. The same results on the M intermediate are observed if deionized blue bR (deI bbR) is treated with these detergents (with or without pH buffers present), even though deionized blue bR containing all the lipids has no photocycle. This suggests that the cation(s) has a role in native bR that is different than in delipidated or monomerized bR, even so far as to suggest that the cation(s) becomes unimportant to the function as the lipids are removed.
Heyes
Colin D
CD
Laser Dynamics Laboratory, School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332-0400, USA.
El-Sayed
Mostafa A
MA
D015502:Absorptiometry, Photon; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000528:radiation effects*; D001692:Biological Transport; D007477:Ions / Q000737:chemistry; D007700:Kinetics; D008055:Lipids / Q000737:chemistry*; D010777:Photochemistry / Q000379:methods; D011487:Protein Conformation / Q000528:radiation effects; D017494:Proton Pumps / Q000737:chemistry / Q000528:radiation effects; D011522:Protons; D013057:Spectrum Analysis
D007477:Ions; D008055:Lipids; D017494:Proton Pumps; D011522:Protons; D001436:Bacteriorhodopsins