Self diffusion and spectral modifications of a membrane protein, the Rubrivivax gelatinosus LH2 complex, incorporated into a monoolein cubic phase.
The light-harvesting complex LH2 from a purple bacterium, Rubrivivax gelatinosus, has been incorporated into the Q230 cubic phase of monoolein. We measured the self-diffusion of LH2 in detergent solution and in the cubic phase by fluorescence recovery after photobleaching. We investigated also the absorption and fluorescence properties of this oligomeric membrane protein in the cubic phase, in comparison with its beta-octyl glucoside solution. In these experiments, native LH2 and LH2 labeled by a fluorescent marker were used. The results indicate that the inclusion of LH2 into the cubic phase induced modifications in the carotenoid and B800 binding sites. Despite these significant perturbations, the protein seems to keep an oligomeric structure. The relevance of these observations for the possible crystallization of this protein in the cubic phase is discussed.
Tsapis
N
N
Laboratoire de Physique Statistique de l'Ecole Normale Supérieure, UMR 8550 CNRS, 75231 Paris Cedex 05, France.
Reiss-Husson
F
F
Ober
R
R
Genest
M
M
Hodges
R S
RS
Urbach
W
W
D004058:Diffusion; D005456:Fluorescent Dyes / Q000378:metabolism; D005989:Glycerides / Q000378:metabolism*; D006217:Halobacterium / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D008823:Micelles; D010777:Photochemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D014961:X-Ray Diffraction
D005456:Fluorescent Dyes; D005989:Glycerides; D008565:Membrane Proteins; D008823:Micelles; D045322:Photosynthetic Reaction Center Complex Proteins; D012996:Solutions; C005953:monoolein