Cytochrome c reductase purified from Crithidia fasciculata contains an atypical cytochrome c1.
Cytochrome c reductase purified from the trypanosomatid Crithidia fasciculata retained antimycin A sensitivity and catalyzed the reduction of horse heart ferricytochrome c in the presence of reduced coenzyme Q10. The complex contained heme b and heme c1 in a ratio of 2:1. Nine major protein bands ranging in size from 55.3 to approximately 12.8 kDa were resolved by SDS-polyacrylamide gel electrophoresis. A 31.6-kDa protein was identified as cytochrome c1 by the presence of a covalently attached heme. A red shift in the alpha-absorbance band of the cytochrome c1 absolute absorbance spectrum, difference absorbance spectrum, and pyridine ferrohemochrome absorbance spectrum suggested that the heme prosthetic group of C. fasciculata cytochrome c1 is bound to the apoprotein through only one thioether bond. A fragment of the cytochrome c1 gene was amplified from C. fasciculata, Trypanosoma brucei, Leishmania tarentolae, and Bodo caudatus. The deduced heme binding site sequence of each of these kinetoplastid species, Phe-Ala-Pro-Cys-His, contains a phenylalanine rather that a cysteine at the first position so that only one thioether bond can be formed between heme and apoprotein. This phenylalanine substitution and the presence of a conserved proline in the sequence may represent compensatory changes that are necessary for optimal interaction of the cytochromes c1 with the atypical cytochromes c of these species.
Priest
J W
JW
Department of Biochemistry, School of Medicine, University of Alabama, Birmingham 35294.
Hajduk
S L
SL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003067:Coenzymes; D016831:Crithidia fasciculata / Q000201:enzymology*; D003574:Cytochrome c Group / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006736:Horses; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology; D009245:NADH Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification*; D010084:Oxidation-Reduction; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000378:metabolism
D003067:Coenzymes; D003574:Cytochrome c Group; D014451:Ubiquinone; D009245:NADH Dehydrogenase; C024989:coenzyme Q10