The general mitochondrial processing peptidase from wheat is integrated into the cytochrome bc1-complex of the respiratory chain.
The bc1-complex (EC 1.10.2.2.) from Triticum aestivum L. was purified by cytochrome-c affinity chromatography and gel filtration using either etiolated seedlings or wheat-germ extract as starting material. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the isolated enzyme revealed ten bands, which were analysed by immunoblotting and direct amino-acid sequencing. The enzyme from wheat is the first bc1-complex that is reported to contain four core proteins (55.5, 55.0, 51.5 and 51.0 kDa). In addition, the wheat bc1-complex comprises cytochrome b (35 kDa), cytochrome c1 (33 kDa) the "Rieske" iron-sulphur protein (25 kDa) and three small subunits < 15 kDa. This composition differs from the one reported in fungi, mammals and potato. Partial sequence determination of the large subunits suggests that the 55.5- and 55.0-kDa-proteins represent the beta-subunit of the general mitochondrial processing peptidase, and the 51.5- and 51.0-kDa proteins the alpha-subunit of this enzyme. The bc1-complex from wheat efficiently processes mitochondrial precursor proteins as shown in an in-vitro processing assay. In control experiments the isolated bc1-complexes from potato, yeast, Neurospora and beef, all purified by the same isolation procedure, were also tested for processing activity. Only the protein complexes from plants contain the general mitochondrial processing peptidase. The composition of the wheat bc1-complex sheds new light on the co-evolution of the processing peptidase and the middle segment of the respiratory chain.
Braun
H P
HP
Institut für Genbiologische Forschung Berlin GmbH, Germany.
Emmermann
M
M
Kruft
V
V
Bödicker
M
M
Schmitz
U K
UK
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004579:Electron Transport; D014450:Electron Transport Complex III / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008666:Metalloendopeptidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D014908:Triticum / Q000201:enzymology*; D000096122:Mitochondrial Processing Peptidase
D014450:Electron Transport Complex III; D008666:Metalloendopeptidases; D009245:NADH Dehydrogenase; D000096122:Mitochondrial Processing Peptidase