Proteins exposed at the surface of chromatophores of Rhodospirillum rubrum: the orientation of isolated chromatophores.
The exposure of proteins at the surface of isolated chromatophores (i.e., the cytoplasmic face of intracytoplasmic membranes) of Rhodospirillum rubrum was studied by proteolysis as well as by enzymatic iodination with 125I. Analyses were performed after polyacrylamide gel electrophoresis of chromatophore proteins solubilized with sodium dodecyl sulfate. Reversible light induced proton uptake by partially digested chromatophores was used as a criterion for the integrity of the permeability barrier and thus, as evidence for proteolysis only of proteins outside of this barrier. Trypsin or alpha-chymotrypsin completely cleaved four proteins which were identified as the heavy subunit of succinate dehydrogenase (Mr = 64 000), the alpha- and beta-subunits of coupling factor ATPase (Mr = 55 000 and 51 000), and the heavy (H) subunit of photochemical reaction centers (Mr = 31 000). alpha-Chymotrypsin, in addition, attacked the protein (Mr = 9000) of light harvesting bacteriochlorophyll preparations. By enzymatic iodination, the same proteins were labeled as were digested with trypsin or alpha-chymotrypsin except for the protein of Mr = 9000. In addition, significant label was incorporated into three more proteins, one of which (Mr = 41 000) could be identified as a major protein of the cell wall. The complete cleavage with trypsin of four proteins exposed at the surface indicated that isolated chromatophores were homogeneously oriented regardless of the method employed for cell breakage, i.e., passage through a French pressure cell at different forces or osmotic shock of sphaeroplasts.
D020130:Bacterial Chromatophores / Q000378:metabolism*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002458:Cell Fractionation / Q000379:methods; D002918:Chymotrypsin; D008027:Light; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D011522:Protons; D012247:Rhodospirillum rubrum / Q000378:metabolism*; D014357:Trypsin
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011522:Protons; D002918:Chymotrypsin; D014357:Trypsin