Reorientations in the bacteriorhodopsin photocycle.
Reversible photoinduced reorientations of bacteriorhodopsin have been detected in suspensions of the purple membrane of Halobacterium salinarium. The anisotropy in bacteriorhodopsin during the nanosecond through millisecond stages of the photocycle was measured by time-resolved linear dichroism and transient absorption measurements. From these measurements the anisotropies of the K, L, M, and O intermediates were determined and related to the chromophore orientation with respect to the initially selected orientation. The anisotropies of the K and L states are 0.38 +/- 0.01 and 0.35 +/- 0.01, respectively. Further anisotropy decay after formation of the M intermediate in about 0.5 ms is evidence of orientational motion at this stage in the photocycle. A constant anisotropy with a value of 0.39 +/- 0.02 in the O intermediate demonstrates a recovery of the initial protein orientation with the formation of the O state. These results demonstrate that reorientations in BR are photoinduced and reversible. Similar measurements for L and M were carried out for purple membrane in polyacrylamide gels, where the anisotropies in the L and M states are 0.38 +/- 0.014 and 0.36 +/- 0.01, respectively. These results show that reorientations also occur in BR immobilized in gels. Anisotropy decay in the M state after formation of the M intermediate was not detected in the gels, in contrast to the M intermediate in suspensions. Orientational changes are observed for BR in purple membrane suspensions in the K state, during the K-->L step, in the M state possibly related to an M1-->M2 transition, and in the O state, where an almost complete return to the original orientation occurs.(ABSTRACT TRUNCATED AT 250 WORDS)
Song
Q
Q
Department of Chemistry, University of Kansas, Lawrence 66045.
Harms
G S
GS
Wan
C
C
Johnson
C K
CK
D000180:Acrylic Resins; D016880:Anisotropy; D001436:Bacteriorhodopsins / Q000737:chemistry*; D006217:Halobacterium / Q000737:chemistry*; D046911:Macromolecular Substances; D010777:Photochemistry; D018717:Purple Membrane / Q000737:chemistry
D000180:Acrylic Resins; D046911:Macromolecular Substances; C016680:polyacrylamide gels; D001436:Bacteriorhodopsins