Time-resolved resonance Raman spectroscopy of bacteriorhodopsin on the millisecond timescale.

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Abstract
A simple technique is described that uses a continuous wave laser with electromechical modulation to obtain time-resolved Raman spectra of transient species on the millisecond timescale. The time behavior of the vibrational bands of the intermediates involved in the proton pumping of bacteriorhodopsin [from Halobacterium halobium] is determined. From these results, along with resonance enhancement and power dependence studies, the bands that appear in the continuous wave Raman spectrum of bacteriorhodopsin can be assigned to 3 intermediates in the photochemical cycle of bacteriorhodopsin, bR570, bL550 and bM412. The Raman spectra of bR570 and bM412 are compared with published spectra of model Schiff bases of all-trans and 13-cis retinal.