Identification and Characterization of an Assembly Intermediate Subcomplex of Photosystem I in the Green Alga Chlamydomonas reinhardtii

• Published in: The Journal of biological chemistry Vol. 285; no. 26; pp. 20072 - 20079
• Main Authors: Ozawa, Shin-ichiro, Onishi, Takahito, Takahashi, Yuichiro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.06.2010; American Society for Biochemistry and Molecular Biology
• Subjects: Algal Proteins - genetics; Algal Proteins - isolation & purification; Algal Proteins - metabolism; Bioenergetics; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - genetics; Chlamydomonas reinhardtii - metabolism; Chlorophyll - isolation & purification; Chlorophyll - metabolism; Chloroplast; Chromatography, Gel; DNA, Algal - genetics; DNA, Chloroplast - genetics; Electron Transfer; Immunoblotting; Light-Harvesting Protein Complexes - genetics; Light-Harvesting Protein Complexes - metabolism; Photosynthesis; Photosystem; Photosystem I Protein Complex - genetics; Photosystem I Protein Complex - isolation & purification; Photosystem I Protein Complex - metabolism; Polymerase Chain Reaction; Protein Assembly; Protein Purification; Reaction Center Complex; Thylakoids - metabolism; Ultracentrifugation; Photosystem; Reaction Center Complex; Electron Transfer; Protein Purification; Photosynthesis; Chloroplast; Protein Assembly
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M109.098954

Photosystem I (PSI) is a multiprotein complex consisting of the PSI core and peripheral light-harvesting complex I (LHCI) that together form the PSI-LHCI supercomplex in algae and higher plants. The supercomplex is synthesized in steps during which 12–15 core and 4–9 LHCI subunits are assembled. Here we report the isolation of a PSI subcomplex that separated on a sucrose density gradient from the thylakoid membranes isolated from logarithmic growth phase cells of the green alga Chlamydomonas reinhardtii. Pulse-chase labeling of total cellular proteins revealed that the subcomplex was synthesized de novo within 1 min and was converted to the mature PSI-LHCI during the 2-h chase period, indicating that the subcomplex was an assembly intermediate. The subcomplex was functional; it photo-oxidized P700 and demonstrated electron transfer activity. The subcomplex lacked PsaK and PsaG, however, and it bound PsaF and PsaJ weakly and was not associated with LHCI. It seemed likely that LHCI had been integrated into the subcomplex unstably and was dissociated during solubilization and/or fractionation. We, thus, infer that PsaK and PsaG stabilize the association between PSI core and LHCI complexes and that PsaK and PsaG bind to the PSI core complex after the integration of LHCI in one of the last steps of PSI complex assembly.