Title

Equilibrium Composition Of Retinal Isomers In Dark-Adapted Bacteriorhodopsin And Effect Of High-Pressure Probed By Near-Infrared Raman-Spectroscopy

Authors

Authors

A. Schulte; L. Bradley;C. Williams

Comments

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Abbreviated Journal Title

Appl. Spectrosc.

Keywords

CHROMOPHORE ISOMERIZATION; PURPLE MEMBRANE; LIGHT ADAPTATION; PROTEIN; VOLUME CHANGE; RAYLEIGH LINE REJECTION; ADAPTATION; PROTEINS; FILTER; EXCITATION; CYCLE; Instruments & Instrumentation; Spectroscopy

Abstract

The combination of near-infrared Raman spectroscopy and variations in external parameters offers new opportunities for site-specific studies of proteins. Using excitation at 840 nm, we have measured the near-infrared Raman spectrum of dark-adapted bacteriorhodopsin at ambient and high pressure. The C=C ethylenic stretching region shows two resolved bands at 1526 and 1534 cm(-1), corresponding to the all-trans and 13-cis isomers. From deconvolution of these bands we find an isomeric ratio between 13-cis and all-trans retinal equal to 1 at ambient pressure. The Raman spectrum gives direct spectroscopic evidence that the 13-cis component is favored at high pressure, implying that it has a smaller volume. The pressure dependence of the isomeric ratio yields a molar volume of -6.6 mL/mol, which suggests ionization of one or two residues or the formation of three hydrogen bonds.

Journal Title

Applied Spectroscopy

Volume

49

Issue/Number

1

Publication Date

1-1-1995

Document Type

Article

Language

English

First Page

80

Last Page

83

WOS Identifier

WOS:A1995QF28300015

ISSN

0003-7028

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