Characterization of PSI isolated from P. chromatophora. (A) Fifteen micrograms of protein of PSI was resolved by SDS/PAGE on an 18% Schägger gel containing 7 M urea. Polypeptides were transferred to PVDF membranes for Western blot analysis using antibodies to various PSI subunits as indicated (lanes 1–5). Additionally, 140 μg protein of isolated PSI was resolved in the same gel system and stained with Sypro Ruby (lane 6); individual polypeptides were excised from the gels for MS analysis (PsaA/PsaB band) or transferred to PVDF membranes for N-terminal sequencing by Edman degradation (PsaE and PsaK1/PsaK2 band). (B) Edman degradation. Sequences shown represent the beginning of the full-length mRNA and their deduced translation products, with predicted initiator methionines encircled and stop codons designated by asterisks. Within the gray box are identities and quantities of the first 10 N-terminal amino acid residues, determined by Edman degradation. (C) Equal concentrations of protein (15 μg per lane) from isolated PSI complexes (left lanes) and from whole-cell extracts (right lanes) were resolved by SDS/PAGE, transferred to PVDF membranes, and subjected to immunoblot analyses using α-PsaC, α-PsaD, α-PsaEpepN, α-PsaF, and α-PsaL. (D) Equal amounts of protein (15 μg per lane) from PSI complexes (lanes 1 and 4), thylakoid membranes (lanes 2 and 5), and whole-cell extracts (lanes 3 and 6) were resolved by SDS/PAGE, stained by CBB (lanes 1–3), or transferred to PVDF membranes and subjected to immunoblot analysis by using α-PsbA, α-PB, and α-PsaEpepN. (lanes 4–6).