Topography of the photosystem I core proteins of the cyanobacterium Synechocystis sp. PCC 6803.
PsaA and PsaB are homologous integral membrane proteins that form the heterodimeric core of photosystem I. Domain-specific antibodies were generated to examine the topography of PsaA and PsaB. The purified photosystem I complexes from the wild type strain of Synechocystis sp. PCC 6803 were treated with eight proteases to study the accessibility of cleavage sites in PsaA and PsaB. Proteolytic fragments were identified using the information from N-terminal amino acid sequencing, reactivity to antibodies, apparent mass, and specificity of proteases. The extramembrane loops of PsaA and PsaB differed in their accessibility to proteases, which indicated the folded structure of the loops or their shielding by the small subunits of photosystem I. NaI-treated and mutant photosystem I complexes were used to identify the extramembrane loops that were exposed in the absence of specific small subunits. The absence of PsaD exposed additional proteolytic sites in PsaB, whereas the absence of PsaE exposed sites in PsaA. These studies distinguish PsaA and PsaB in the structural model for photosystem I that has been proposed on the basis of x-ray diffraction studies (Krauss, N., Schubert, W.-D., Klukas, O., Fromme, P., Witt, H. T., and Saenger, W. (1996) Nat. Struct. Biol. 3, 965-973). Using osmotically shocked cells for protease treatments, the N terminus of PsaA was determined to be on the n side of the photosynthetic membranes. Based on these data and available published information, we propose a topological model for PsaA and PsaB.
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Department of Biochemistry and Biophysics, Iowa State University, Ames, Iowa 50011, USA.
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D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002417:Cattle; D018360:Crystallography, X-Ray; D000458:Cyanobacteria / Q000737:chemistry*; D010450:Endopeptidases / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics; D045331:Photosystem I Protein Complex; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation; D017510:Protein Folding; D013552:Swine
D001426:Bacterial Proteins; D008565:Membrane Proteins; C073973:PSI-A protein, Synechococcus; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D010940:Plant Proteins; C070290:PsaD protein, cyanobacteria; C089909:photosystem I, psaB subunit; C079741:photosystem I, psaE subunit; C079742:psaF protein, bacteria; D010450:Endopeptidases