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Phosphorus 31 nmr study of the interaction of cations with purple membrane and of the purple blue transition

Phosphorus 31 nmr study of the interaction of cations with purple membrane and of the purple blue transition

Biochemistry 27(18): 7009-7015

ISSN/ISBN: 0006-2960

DOI: 10.1021/bi00418a051

We have studied by 31P NMR both the native purple membrane and the deionized membrane obtained by removal of endogenous cations. The latter membrane was shown to undergo a blue to purple color transition with increasing pH (apparent pK = 5.6). In parallel with this color transition, the 31P NMR spectrum obtained at high membrane concentration was dramatically changed from a normal bilayer powder pattern to a seemingly "inverted" line shape. This effect was not observed in native purple membrane and was found to be reversed by addition of cations to the deionized purple membrane. Several data indicated that this inversion of the 31P NMR line shape is due to orientation of the membrane fragments perpendicular to the magnetic field. Further studies indicated that both native and deionized purple membranes can undergo such magnetic orientation but that the favorable concentration range is greatly increased for the deionized preparation. This effect is attributed to differences in bacteriorhodopsin conformation and/or membrane surface charge in the two membranes. Binding of divalent cations to the purple membrane was shown to promote an increase of the chemical shift anisotropy of phospholipid phosphate groups as revealed by 31P NMR. Accordingly, binding of a trivalent paramagnetic cation promoted strong broadening of the 31P NMR spectrum. This suggests a close spatial or structural relationship between phospholipid head groups and cation binding sites in the purple membrane.

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Accession: 006106467

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