Purification and characterization of Mycobacterium tuberculosis 1D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase, MshB, a mycothiol biosynthetic enzyme

Protein Expression and Purification
Gerald L NewtonRobert C Fahey

Abstract

Mycothiol (MSH, AcCys-GlcN-Ins) is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis. MshB, the GlcNAc-Ins deacetylase, is a key enzyme in MSH biosynthesis. MshB from M. tuberculosis was cloned, expressed, purified, and its properties characterized. Values of k(cat) and K(m) for MshB were determined for the biological substrate, GlcNAc-Ins, and several other good substrates. The substrate specificity of MshB was compared to that of M. tuberculosis mycothiol S-conjugate amidase (Mca), a homologous enzyme having weak GlcNAc-Ins deacetylase activity. Both enzymes are metalloamidases with overlapping amidase activity toward mycothiol S-conjugates (AcCySR-GlcN-Ins). The Ins residue and hydrophobic R groups enhance the activity with both MshB and Mca, but changes in the acyl group attached to GlcN have opposite effects on the two enzymes.

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Citations

Oct 30, 2010·The Journal of Biological Chemistry·Henrieta SkovierováMary Jackson
Oct 24, 2012·The Journal of Biological Chemistry·Arek K RainczukPaul K Crellin
Sep 6, 2008·Microbiology and Molecular Biology Reviews : MMBR·Gerald L NewtonRobert C Fahey
Jun 28, 2014·Biomolecules·Shane ViarsMarcy Hernick
Jul 18, 2015·Expert Review of Anti-infective Therapy·Varahenage R PereraKit Pogliano
Aug 1, 2008·Expert Review of Respiratory Medicine·Haruaki TomiokaToshiaki Shimizu
Jun 6, 2015·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Evren KocabasMarcy Hernick
Jul 28, 2016·Journal of Structural Biology·Tsutomu NakamuraKoichi Uegaki
Sep 11, 2007·Biochemical and Biophysical Research Communications·Mamta RawatYossef Av-Gay
Nov 26, 2008·Natural Product Reports·Vishnu Karthik Jothivasan, Chris J Hamilton
Feb 9, 2007·FEMS Microbiology Reviews·Mamta Rawat, Yossef Av-Gay
Aug 31, 2006·The Journal of Biological Chemistry·Gerald L NewtonRobert C Fahey
Jun 25, 2015·Microbiology Spectrum·Bridgette M CummingAdrie J C Steyn
Oct 19, 2012·Biochimica Et Biophysica Acta·Robert C Fahey

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