Identification of 42 possible cytochrome C genes in the Shewanella oneidensis genome and characterization of six soluble cytochromes.
Through pattern matching of the cytochrome c heme-binding site (CXXCH) against the genome sequence of Shewanella oneidensis MR-1, we identified 42 possible cytochrome c genes (27 of which should be soluble) out of a total of 4758. However, we found only six soluble cytochromes c in extracts of S. oneidensis grown under several different conditions: (1) a small tetraheme cytochrome c, (2) a tetraheme flavocytochrome c-fumarate reductase, (3) a diheme cytochrome c4, (4) a monoheme cytochrome c5, (5) a monoheme cytochrome c', and (6) a diheme bacterial cytochrome c peroxidase. These cytochromes were identified either through N-terminal or complete amino acid sequence determination combined with mass spectroscopy. All six cytochromes were about 10-fold more abundant when cells were grown at low than at high aeration, whereas the flavocytochrome c-fumarate reductase was specifically induced by anaerobic growth on fumarate. When adjusted for the different heme content, the monoheme cytochrome c5 is as abundant as are the small tetraheme cytochrome and the tetraheme fumarate reductase. Published results on regulation of cytochromes from DNA microarrays and 2D-PAGE differ somewhat from our results, emphasizing the importance of multifaceted analyses in proteomics.
Meyer
Terry E
TE
Department of Biochemistry and Molecular Biophysics University of Arizona, Tucson, Arizona 85721, USA. temeyer@u.arizona.edu
Tsapin
Alexandre I
AI
Vandenberghe
Isabel
I
de Smet
Lina
L
Frishman
Dmitrij
D
Nealson
Kenneth H
KH
Cusanovich
Michael A
MA
van Beeumen
Jozef J
JJ
D000595:Amino Acid Sequence; D002455:Cell Division; D003574:Cytochrome c Group / Q000737:chemistry; D003580:Cytochromes / Q000378:metabolism; D045304:Cytochromes c / Q000235:genetics* / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D005650:Fumarates / Q000737:chemistry; D016680:Genome, Bacterial; D006418:Heme / Q000737:chemistry; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D020543:Proteome; D017386:Sequence Homology, Amino Acid; D020592:Shewanella / Q000235:genetics*; D013053:Spectrophotometry; D014466:Ultraviolet Rays
D003574:Cytochrome c Group; D003580:Cytochromes; D005650:Fumarates; D020543:Proteome; D006418:Heme; C046450:cytochrome C5; C032005:fumaric acid; D045304:Cytochromes c