Molecular cloning, expression, and functional analysis of a predicted sulfotransferase STF9 from Mycobacterium avium

Md Murad Hossain, Yuuji Moriizumi, Shotaro Tanaka, Makoto Kimura, Yoshimitsu Kakuta

研究成果: Contribution to journalArticle査読

3 被引用数 (Scopus)

抄録

Sulfotransferases catalyze the transfer of sulfate group from para-nitrophenyl sulfate (pNPS) or 3′-phosphoadenosine-5′- phosphosulfate (PAPS) onto acceptor molecules in the biosynthesis of sulfate esters. Human pathogenic mycobacteria are known to produce numerous sulfated molecules on their cell surface which have been implicated as important mediators in host-pathogen interactions. The open reading frame stf9, a predicted homologue of sulfotransferase in the Mycobacterium avium genomic data, was cloned and over expressed in Escherichia coli. The recombinant STF9 conserved the characteristic PAPS binding motif of sulfotransferase and was purified as a 44 kDa soluble protein which exhibited transfer of sulfate group from pNPS (K m 1.34 mM, V max 7.56 nmol/min/mg) onto 3′-phosphoadenosine-5′-phosphate (K m 0.24 mM, V max 10.36 nmol/min/mg). The recombinant STF9 protein was also capable of transferring sulfate group from PAPS onto certain acceptor substrates in E. coli, and showed binding affinity to the PAP-agarose resin, supporting the sulfotransferase activity of the recombinant STF9 protein. This is the first report of molecular evidence for sulfotransferase activity of a protein from M. avium. Mutation of Arg96 to Ala and Glu170 to Ala abolishes sulfotransferase activity, indicating the importance of Arg96 and Glu170 in STF9 activity catalysis.

本文言語英語
ページ(範囲)155-162
ページ数8
ジャーナルMolecular and cellular biochemistry
350
1-2
DOI
出版ステータス出版済み - 4 1 2011

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

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