GltX from Clostridium saccharobutylicum NCP262: glutamate synthase or oxidoreductase?
A full-length gene encoding a homologue of the small subunit of the glutamate synthase (GOGAT) enzyme was isolated from the anaerobic bacterium, Clostridium saccharobutylicum NCP262, which has been used extensively for the commercial production of solvents. Using a screening system designed to isolate genes involved in electron transport, plasmid pMET13C1 was isolated. Analysis of this plasmid identified a gene (1245 bp) with a predicted approximately 46-kDa product, which was associated with reductive activation of the pro-drug metronidazole. The deduced 414-amino-acid sequence was not typical of electron transport proteins, but rather shared striking homology to the small (beta) subunit of the GOGAT enzyme and other beta subunit-like polypeptides, and was thus designated gltX. Although all the functional domains typical of GOGAT beta subunits were conserved in this GltX protein, certain sequence features were not conserved. Furthermore, it was independently transcribed, did not lie adjacent to a GOGAT large subunit (alpha) domain, and its expression was not regulated by nitrogen conditions. These results provide additional support for current theories on the evolutionary relationships of GOGAT beta subunit domains in bacteria, and suggest that GltX belongs to a more general family of oxidoreductases, which is used in a context other than glutamate biosynthesis to transfer electrons to a currently unknown protein domain.
Stutz
Helen E
HE
Department of Molecular and Cell Biology, University of Cape Town, Private Bag Rondebosch 7701, South Africa.
Reid
Sharon J
SJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003013:Clostridium / Q000201:enzymology* / Q000235:genetics*; D019143:Evolution, Molecular; D040461:Gene Components; D005970:Glutamate Synthase / Q000235:genetics*; D008795:Metronidazole / Q000378:metabolism; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013053:Spectrophotometry
D008795:Metronidazole; D010088:Oxidoreductases; D005970:Glutamate Synthase