Membrane topography of the photosynthetic reaction center polypeptides of Rhodopseudomonas sphaeroides.
The topography of the photosynthetic reaction center (RC) polypeptides (H, M, and L) was investigated by proteolysis and radioiodination of membrane vesicles isolated from Rhodopseudomonas sphaeroides. Chromatophores, obtained from French-pressed cell lysates, are closed vesicles' and oriented inside out with respect to the cytoplasmic membrane (cytoplasmic side out). Spheroplast-derived vesicles (SDVs), obtained after osmotic lysis of lysozyme-treated cells, are oriented right side in (periplasmic side out). Alpha-Chymotrypsin treatment of chromatophores and trypsin treatment of SDVs resulted in cleavage of H. Alpha-Chymotrypsin treatment of SDVs did not cleave H, and trypsin treatment of chromatophores did not consistently cleave this polypeptide. M and L of both vesicles were apparently not affected by these proteases. The SDV trypsin cleavage product of H was identified by alpha-chymotryptic (125)I-labeled peptide mapping and had a molecular weight of 26 000. Membrane surface radioiodination with chloroglycoluril coated on glass tubes resulted in preferential labeling of H and M of SDVs and chromatophores. The radiospecific activities of H, M, and L were higher with labeling of SDVs as compared to labeling of chromatophores. Alpha-Chymotryptic (125)I-labeled peptide maps of H, M, and L from surface-radioiodinated SDVs differed from the corresponding maps of these polypeptides from surface-radioiodinated chromatophores. The results indicate the asymmetric exposure of H, M, and L on opposite surfaces of the R. sphaeroides membrane. Exposed iodination sites of these polypeptides are more abundant on the periplasmic surface than on the cytoplasmic surface of this membrane.
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002918:Chymotrypsin; D007700:Kinetics; D010446:Peptide Fragments / Q000032:analysis; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins; D011487:Protein Conformation; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D014357:Trypsin
D001426:Bacterial Proteins; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D002918:Chymotrypsin; D014357:Trypsin