Time-resolved studies of the protein bacteriorhodopsin using femtosecond laser pulses.

摘要

The primary events in the all-trans to 13-cis photoisomerization of retinal in bacteriorhodopsin have been investigated with femtosecond time-resolved absorbance spectroscopy. Spectra measured over a broad range extending from 7,000-22,400 cm{dollar}sp{lcub}-1{rcub}{dollar} reveal features whose dynamics are inconsistent with a model proposed earlier to account for the highly efficient photoisomerization process. Emerging from this work is a new three-state model. Photoexcitation of retinal with visible light accesses a shallow well on the excited state potential energy surface. This well is bounded by a small barrier, arising from an avoided crossing that separates the Franck-Condon region from the nearby reactive region of the photoisomerization coordinate. At ambient temperatures, the reactive region is accessed with a time constant of {dollar}sim{dollar}500 fs, whereupon the retinal rapidly twists and encounters a second avoided crossing region. The driving force for photoisomerization resides in the retinal, not in the surrounding protein. This view contrasts with an earlier model where photoexcitation was thought to access directly a reactive region of the excited state potential and thereby drive the retinal to a twisted conformation within 100-200 fs.