X-ray Structure of Bacteriorhodopsin at 2.5 Angstroms from Microcrystals Grown in Lipidic Cubic Phases
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; A. E. Blaurock ibid. p. 139.
Henderson R., et al., ibid. 213, 899 (1990).
Engelman D. M., Henderson R., McLachlan A. D., Wallace B. A., Proc. Natl. Acad. Sci. U.S.A. 77, 2023 (1980).
Ovchinnikov Y. A., Abdulaev N. G., Feigina M. Y., Kiselev A. V., Lobanov N. A., FEBS Lett. 100, 219 (1979);
Krebs M. P., Hauss T., Heyn M. P., Rajbhandary U. L., Khorana H. G., Proc. Natl. Acad. Sci. U.S.A. 88, 859 (1991);
. Monoolein (1-monooleoyl- rac -glycerol C 18:1c9 ) was obtained from Sigma.
At room temperature a powder pattern with one strong diffraction ring at 53 Å (accessible low-resolution limit is 60 Å) was observed. Upon rapid cooling to liquid nitrogen temperature the diffraction image changed to multiple low-resolution rings at 52 Å 25 Å 12 Å and a strong ring at 4.5 Å. The latter is also observed at 4°C and can be assigned to the periodicity of the lipid chain packing [
]. This transition also manifests itself by the fact that the cubic phase optically transparent at room temperature turns slightly opaque upon cooling.
The rms deviations are 0.7 and 1.2 Å for the main chain and for all the non-hydrogen atoms respectively if only the helices are considered. This compares with rms deviations of 3.6 Å (main chain) and 4.0 Å (all non-hydrogen atoms) if all residues between 7 and 225 are included.
The degree of distortion was unclear. A recent nuclear magnetic resonance study claims that the angles for the C18 C19 and C20 methyl groups with the membrane normal are 37° 40° and 32° respectively [
] whereas the EM structure represents them as 38.6° 29.3° and 16.1° respectively. Our structure reveals values of 32° 34° and 10° respectively.
J. Heberle and E. M. Landau unpublished results.
Z. Otwinowski in Data Collection and Processing L. Sawyer N. Isaacs S. Bailey Eds. (SERC Daresbury Laboratory Warrington UK 1993) pp. 56–62.
Jones A. T., Zou J. Y., Cowan S. W., Kjeldgaard M., ibid. A47, 110 (1991).
A. T. Brünger X-PLOR Version 3.1: A System for X-ray Crystallography and NMR (Yale Univ. Press New Haven CT 1992).
We thank C. Riekel H. Belrahli and A. Bram for invaluable help on the ID13 beamline; E. Fanchon R. Kahn and M. Roth on the French beamline D2AM; G. Büldt for his generosity; G. Büldt S. Cusack O. Dideberg J. Heberle J. Sass and G. Zaccai for stimulating discussions; and T. Bickle and R. Eisenberg for critically reading the manuscript. Supported by the Université Joseph Fourier the Institut de Biologie Structurale the Institut Universitaire de France the University of Basel and grants from the Swiss National Science Foundation (SPP 5002-37911 and -46092) EU-BIOMED (BIO4CT96/BBW95.0137.2) and EU-BIOTECH (EU-BIO4CT96/BBW96.0145.1). Beam time at the ESRF (Grenoble France) was granted under experiments LS-435 LS-553 LS-655 02.03.060 and 02.03.111.
