Interconversions among four M-intermediates in the bacteriorhodopsin photocycle.
Halobacterium salinarum displays four distinct kinetic forms of M-intermediate in its bacteriorhodopsin photocycle. In wild-type, there are mainly two species with time constants near 2 and 5 ms. Under various kinds of stress, two other species arise with time constants near 10 and 70 ms. We show that these four species are interconvertible. Increases in membrane hydrophobicity convert the slower to faster forms. Perturbations caused by Triton X-100 or mutations convert faster to slower forms. The fastest form requires a hydrophobic membrane environment near a ring of four charged aspartate residues in the trimer, namely Asp36, Asp38, Asp102, and Asp104 in the cytoplasmic loop regions. Interconversions of the 2-ms and 5-ms species of the wild-type are accomplished by pH-changes. The potential significance of these findings is discussed.
Hendler
Richard W
RW
Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-0510, USA. rwh@helix.nih.gov
Bose
Salil
S
D000473:Alkanes / Q000737:chemistry; D019943:Amino Acid Substitution; D001224:Aspartic Acid / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019617:Halobacterium salinarum / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008027:Light; D008563:Membrane Lipids / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D011522:Protons; D013053:Spectrophotometry / Q000379:methods / Q000706:statistics & numerical data
D000473:Alkanes; D001426:Bacterial Proteins; D008563:Membrane Lipids; D011522:Protons; D001224:Aspartic Acid; D001436:Bacteriorhodopsins