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Effect of fluorescamine modification of purple membranes on exciton coupling and light-to-dark adaptation
Authors:Eric Lam  Lester Packer
Affiliation:1. Membrane Bioenergetics Group, Lawrence Berkeley Laboratory, University of California, Berkeley, California 94720 USA.;2. Department of Physiology-Anatomy University of California, Berkeley, California 94720 USA.;3. Department of Biophysics, University of California, Berkeley, California 94720 USA.
Abstract:Purple membranes of Halobacterium, halobium were modified with fluorescamine. At pH 8.8, with a molar ratio of fluorescamine to bacteriorhodopsin of 170, about 6 residues of lysine were modified while the arginines were not affected at all. Except for the appearance of the fluorescamine peak at 394 nm and some broadening of the chromophore peak at 570 nm, the absorption spectrum of bacteriorhodopsin was not significantly changed after modification. After fluorescamine modification, circular dichroism studies indicated loss of exciton coupling between bacteriorhodopsin molecules in the purple membrane. Rotational diffusion studies suggested enhanced mobility of the chromophore after modification. However, the spectral changes accompanying the light-to-dark adaptation of purple membranes were not prevented by fluorescamine modification. The implications of these findings are that exciton coupling between neighboring bacteriorhodopsin molecules in the purple membrane is not required for light-to-dark adaptation.
Keywords:HEPES  N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid  BR  bacteriorhodopsin  EA  ethyl acetimidate
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