Characterization of a new type of sulfite dehydrogenase from Paracoccus pantotrophus GB17

Item Type:	Article
Title:	Characterization of a new type of sulfite dehydrogenase from Paracoccus pantotrophus GB17
Creators Name:	Quentmeier, A. and Kraft, R. and Kostka, S. and Klockenkaemper, R. and Friedrich, C.G.
Abstract:	The periplasmic sulfite dehydrogenase of Paracoccus pantotrophus GB17 was purified to homogeneity by a four-step procedure from cells grown lithoautotrophically with thiosulfate. The molecular mass of native sulfite dehydrogenase was 190 kDa as determined by native gradient PAGE. SDS-PAGE showed sulfite dehydrogenase to comprise two subunits with molecular masses of 47 kDa and 50 kDa, suggesting an {alpha}2{beta}2 structure. The N-terminal amino acid sequence and immunochemical analysis using SoxC-specific antibodies identified the 47-kDa protein as the soxC gene product. SoxD-specific antibodies identified the 50-kDa protein as SoxD. Based on the molecular masses deduced from the nucleotide sequence for mature SoxC (43,442 Da) and SoxD (37,637 Da) sulfite dehydrogenase contained 1.30 mol molybdenum/mol {alpha}2{beta}2 sulfite dehydrogenase. The iron content was 3.17 mol/mol α2β2 sulfite dehydrogenase, and 3.53 mol heme/mol {alpha}2{beta}2 sulfite dehydrogenase was determined by pyridine hemochrome analysis. These data are consistent with the two heme-binding domains (CxxCH), characteristic for c-type cytochromes, deduced from the soxD nucleotide sequence. Electrospray ionization revealed two masses for SoxC of 43,503 and 43,897 Da. The difference in molecular mass was attributed to the molybdenum cofactor of SoxC. For SoxD a mass of 38,815 Da was determined; this accounted for the polypeptide and two covalently bound hemes. Reconstitution of the catalytic activity of sulfite dehydrogenase required additional fractions; these eluted from Q Sepharose at 0.05, 0.25, and 0.30 M NaCl. The K(m) of sulfite dehydrogenase for sulfite was 7.0 {mu}M and for cytochrome c 19 {mu}M. Sulfite dehydrogenase activity was inhibited by sulfate and phosphate. The structural and catalytic properties make sulfite dehydrogenase from P. denitrificans GB17 distinct from sulfite oxidases of other prokaryotic or eukaryotic sources.
Keywords:	Diheme Cytochrome c, Molybdenum Cofactor, Paracoccus Denitrificans, Paracoccus Pantotrophus, Periplasmic Sulfite Dehydrogenase, Thiosulfate Oxidation
Source:	Archives of Microbiology
ISSN:	0302-8933
Publisher:	Springer
Volume:	173
Number:	2
Page Range:	117-125
Date:	1 February 2000
Official Publication:	https://doi.org/10.1007/s002039900118
PubMed:	View item in PubMed

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