Location and orientation of the chromophore in bacteriorhodopsin. Analysis by fluorescence energy transfer

Tsutomu Kouyama, Yoshiaki Kimura, Kazuhiko Kinosita, Akira Ikegami

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The spatial location and orientation of the retinal chromophore in bacteriorhodopsin were estimated from a fluorescence energy transfer study. The energy donor used in this study was a fluorescent retinal derivative, which was obtained by partial reduction of the purple membrane with sodium borohydride, and the energy acceptor was the native chromophore remaining in the same membrane. Since bacteriorhodopsin forms a two-dimensional crystal with P3 symmetry in the purple membrane, and the membrane structure is maintained after the reduction, the rate of energy transfer from a donor to any acceptor existing in the same membrane can be calculated as a function of the location and orientation of the chromophores in the unit cell. Quantitative analyses of the fluorescence decay curve and the quantum yield, with various extents of reduction, enabled us to determine the most probable location and orientation. The result suggested that the chromophore was situated near the centre of the protein in such an orientation that the dipole-dipole interaction with neighbouring chromophores was close to minimum.

Original languageEnglish
Pages (from-to)337-359
Number of pages23
JournalJournal of Molecular Biology
Volume153
Issue number2
DOIs
Publication statusPublished - 1981 Dec 5
Externally publishedYes

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Bacteriorhodopsins
Energy Transfer
Purple Membrane
Fluorescence
Membranes
Proteins

ASJC Scopus subject areas

  • Virology

Cite this

Location and orientation of the chromophore in bacteriorhodopsin. Analysis by fluorescence energy transfer. / Kouyama, Tsutomu; Kimura, Yoshiaki; Kinosita, Kazuhiko; Ikegami, Akira.

In: Journal of Molecular Biology, Vol. 153, No. 2, 05.12.1981, p. 337-359.

Research output: Contribution to journalArticle

Kouyama, Tsutomu ; Kimura, Yoshiaki ; Kinosita, Kazuhiko ; Ikegami, Akira. / Location and orientation of the chromophore in bacteriorhodopsin. Analysis by fluorescence energy transfer. In: Journal of Molecular Biology. 1981 ; Vol. 153, No. 2. pp. 337-359.
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