Photochemistry in dried polymer films incorporating the deionized blue membrane form of bacteriorhodopsin

Biophys J. 1998 Oct;75(4):1619-34. doi: 10.1016/S0006-3495(98)77605-2.

Abstract

The preparation and photochemical properties of dried deionized blue membrane (dIbR600; lambdamax approximately 600 nm, epsilon approximately 54, 760 cm-1 M-1, f approximately 1.1) in polyvinyl alcohol films are studied. Reversible photoconversion from dIbR600 to the pink membrane (dIbR485; lambdamax approximately 485 nm) is shown to occur in these films under conditions of strong 647-nm laser irradiation. The pink membrane analog, dIbR485, has a molar extinction coefficient of approximately 39,000 cm-1 M-1 (f approximately 1.2). The ratio of pink --> blue and blue --> pink quantum efficiencies is 33 +/- 5. We observe an additional blue-shifted species (dIbR455, lambdamax approximately 455 nm) with a very low oscillator strength (f approximately 0.6, epsilon approximately 26,000 cm-1 M-1). This species is the product of fast thermal decay of dIbR485. Molecular modeling indicates that charge/charge and charge/dipole interactions introduced by the protonation of ASP85 are responsible for lowering the excited-state all-trans --> 9-cis barrier to approximately 6 kcal mol-1 while increasing the corresponding all-trans --> 13-cis barrier to approximately 4 kcal mol-1. Photochemical formation of both 9-cis and 13-cis photoproducts are now competitive, as is observed experimentally. We suggest that dIbR455 may be a 9-cis, 10-s-distorted species that partially divides the chromophore into two localized conjugated segments with a concomitant blue shift and decreased oscillator strength of the lambdamax absorption band.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Archaeal Proteins*
  • Bacteriorhodopsins / chemistry*
  • Bacteriorhodopsins / radiation effects
  • Binding Sites
  • Carotenoids*
  • Halorhodopsins*
  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Photochemistry / methods
  • Protein Conformation*
  • Quantum Theory
  • Sensory Rhodopsins*
  • Spectrophotometry / instrumentation
  • Spectrophotometry / methods

Substances

  • Archaeal Proteins
  • Halorhodopsins
  • Sensory Rhodopsins
  • sensory rhodopsin II protein, archaeal
  • Carotenoids
  • Bacteriorhodopsins