Effect of partial delipidation of purple membrane on the photodynamics of bacteriorhodopsin.
The effect of lipid-protein interaction on the photodynamics of bacteriorhodopsin (bR) was investigated by using partially delipidated purple membrane (pm). When pm was incubated with a mild detergent, Tween 20, the two major lipid components of pm, phospholipids and glycolipids, were released in different ways: the amount of phospholipids released was proportional to the logarithm of the incubation time; the release of glycolipids became noticeable after the release of approximately 2 phospholipids/bR, but soon leveled off at approximately 50% of the initial content. It was found that the thermal decay of the photocycle intermediate N560 was inhibited by the removal of less than 2 phospholipids per bR. This inhibition was partly explained by an increase in the local pH near the membrane surface. More significant changes in the bR photoreactions were observed when greater than 2 phospholipids/bR were removed: (1) the extent of light adaptation became much smaller, and this reduction correlated with the release of glycolipids; (2) N560 became difficult to detect; (3) the M412 intermediate, which is characterized by a pH-insensitive lifetime, was replaced by a long-lived M-like photoproduct with a pH-sensitive lifetime. The heavy delipidation apparently altered the mechanism by which the deprotonated Schiff base receives a proton. An important conformational change in the protein moiety is suggested to take place during the M412 state, this conformational change being inhibited in the rigid lipid environment.
Fukuda
K
K
Institute of Physical and Chemical Research, Saitama, Japan.
Ikegami
A
A
Nasuda-Kouyama
A
A
Kouyama
T
T
D001436:Bacteriorhodopsins / Q000032:analysis*; D002462:Cell Membrane / Q000032:analysis; D006017:Glycolipids / Q000302:isolation & purification; D006217:Halobacterium / Q000032:analysis*; D007536:Isomerism; D007700:Kinetics; D008027:Light; D008055:Lipids / Q000302:isolation & purification*; D010743:Phospholipids / Q000302:isolation & purification*; D011136:Polysorbates
D006017:Glycolipids; D008055:Lipids; D010743:Phospholipids; D011136:Polysorbates; D001436:Bacteriorhodopsins