Reversal of the surface charge asymmetry in purple membrane due to single amino acid substitutions.

Abstract:

:Twenty-seven mutant bacteriorhodopsin's were screened to determine the PKa for reversal of the permanent electric dipole moment. The photoelectric response of an aqueous purple-membrane suspension was used to determine the direction of the purple-membrane dipole moment as a function of pH. The pK(a) for the dipole reversal of wild-type bacteriorhodopsin is 4.5. Six of the 27 mutant bacteriorhodopsin's were found to have a pK(a) for dipole reversal larger than that of wild-type bacteriorhodopsin. Two of these mutants, L93T and L93W, involve a neutral amino acid substitution in the interior of the protein. The direction of the purple-membrane permanent electric dipole moment is determined by the purple-membrane surface charge asymmetry. We conclude that these two substitutions, which do not involve charge replacement, alter the pK(a) for the reversal of the purple-membrane surface charge asymmetry. We suggest that these changes to the pK(a) are due to altered protein folding at the surface of the purple-membrane induced by single-site substitutions in the protein interior.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Hsu KC,Rayfield GW,Needleman R

doi

10.1016/S0006-3495(96)79802-8

subject

Has Abstract

pub_date

1996-05-01 00:00:00

pages

2358-65

issue

5

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(96)79802-8

journal_volume

70

pub_type

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