Identification of mitochondrial proteins in membrane preparations from Chlamydomonas reinhardtii.
Preparations enriched in Chlamydomonas reinhardtii thylakoids have proven useful in the study of photosynthesis. Many of their polypeptides however remain unidentified. We report here on three of those, h1 (34 kDa), h2 (11 kDa), and P3 (63 kDa). h1, h2, and P3 are present in all tested mutants of C. reinhardtii lacking either one or several of the photosynthetic chain complexes or depleted in thylakoid membranes. h2 is an ascorbate-reducible, soluble c550-type cytochrome encoded in the nucleus. It cross-reacts immunologically with mitochondrial cytochromes c from various sources and contains a hexapeptide encoded in C. reinhardtii cytochrome c cDNA. P3, a nuclear-encoded peripheral protein, cross-reacts with various ATP synthase beta subunits. Its N-terminal sequence is encoded in C. reinhardtii mitochondrial beta subunit cDNA. h1 behaves as an integral hemoprotein; it is absent in a mitochondrial mutant that carries a deletion in apocytochrome b gene. We conclude that C. reinhardtii mitochondrial membranes copurify with thylakoid membranes. h1 is part of the cytochrome bc1 complex, h2 is cytochrome c, and P3 is the beta subunit of mitochondrial ATP synthase.
Atteia
A
A
Institut de Biologie Physico-Chimique, Centre National de la Recherche Scientifique URA1187, Paris, France.
de Vitry
C
C
Pierre
Y
Y
Popot
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000201:enzymology*; D002736:Chloroplasts / Q000201:enzymology; D003429:Cross Reactions; D003574:Cytochrome c Group / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000032:analysis*; D006420:Hemeproteins / Q000032:analysis; D007425:Intracellular Membranes / Q000201:enzymology; D008565:Membrane Proteins / Q000032:analysis*; D008928:Mitochondria / Q000737:chemistry / Q000201:enzymology*; D008969:Molecular Sequence Data; D010788:Photosynthesis; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013057:Spectrum Analysis
D003574:Cytochrome c Group; D005656:Fungal Proteins; D006420:Hemeproteins; D008565:Membrane Proteins; D006180:Proton-Translocating ATPases