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: ジャーナルへの寄稿記事

3 引用 (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.

元の言語英語
ページ(範囲)155-162
ページ数8
ジャーナルMolecular and cellular biochemistry
350
発行部数1-2
DOI
出版物ステータス出版済み - 4 1 2011

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Sulfotransferases
Mycobacterium avium
Functional analysis
Cloning
Molecular Cloning
Phosphoadenosine Phosphosulfate
Sulfates
Recombinant Proteins
Escherichia coli
Host-Pathogen Interactions
Molecules
Biosynthesis
Pathogens
Mycobacterium
Catalysis
Sepharose
Open Reading Frames
Esters
Proteins
Resins

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Molecular cloning, expression, and functional analysis of a predicted sulfotransferase STF9 from Mycobacterium avium. / Hossain, Md Murad; Moriizumi, Yuuji; Tanaka, Shotaro; Kimura, Makoto; Kakuta, Yoshimitsu.

:: Molecular and cellular biochemistry, 巻 350, 番号 1-2, 01.04.2011, p. 155-162.

研究成果: ジャーナルへの寄稿記事

Hossain, Md Murad ; Moriizumi, Yuuji ; Tanaka, Shotaro ; Kimura, Makoto ; Kakuta, Yoshimitsu. / Molecular cloning, expression, and functional analysis of a predicted sulfotransferase STF9 from Mycobacterium avium. :: Molecular and cellular biochemistry. 2011 ; 巻 350, 番号 1-2. pp. 155-162.
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