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

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Abstract

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.

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Acknowledgment

This study was supported by a grant from the National Project on Protein Structural and Functional Analyses from the Ministry of Education, Culture, Sports, Science, and Technology, Japan.

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  1. Md. Murad Hossain

    Present address: Department of Cell Biology, Institute of Nephrology, Niigata University, 1-757 Asahimachi-Dori, Niigata, 951-8510, Japan

Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

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

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  1. Md. Murad Hossain
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  2. Yuuji Moriizumi
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  3. Shotaro Tanaka
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  4. Makoto Kimura
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  5. Yoshimitsu Kakuta
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Correspondence to Md. Murad Hossain.

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Hossain, M.M., Moriizumi, Y., Tanaka, S. et al. Molecular cloning, expression, and functional analysis of a predicted sulfotransferase STF9 from Mycobacterium avium . Mol Cell Biochem 350, 155–162 (2011). https://doi.org/10.1007/s11010-010-0693-1

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  • DOI: https://doi.org/10.1007/s11010-010-0693-1

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