Please use this identifier to cite or link to this item: http://nopr.niscair.res.in/handle/123456789/9809
Title: Mutational analysis for enzyme activity of mouse Gal1,3GalNAc 2,3-sialyltransferase (mST3Gal I)
Authors: Kim, Kyoung-Sook
Oh, Seoung-Un
Lee, Jai-Heon
Lee, Young-Choon
Keywords: Sialyltransferase
Chemical modification
Site-directed mutagenesis
Issue Date: Jun-2010
Publisher: CSIR
Abstract: To determine which amino acid residues are essential for the catalytic activity of mouse Gal1,3GalNAc 2,3-sialyltransferase (mST3Gal I), chemical modification and site-directed mutagenesis were employed against tryptophan and cysteine residues located in the predicted catalytic domain. This enzyme was strongly inhibited by N-bromosuccinimide, a specific blocking reagent for tryptophan residues, and the enzyme activity was completely lost at 3 mM, suggesting the involvement of tryptophan residues in the catalytic activity of mST3Gal I. The N-ethylmaleimide, an irreversible reagent for sulfhydryl group, significantly inhibited the enzyme activity. Seven tryptophan and six cysteine residues conserved in the cloned Gal1,3GalNAc 2,3-sialyltransferases were separately substituted into phenylalanine and serine, respectively. The enzymatic activity assay for tryptophan mutants produced in COS cells showed a complete abolishment of the activity in all of the mutants, except that W70F and W97F retained about 60% and 40% activities of wild type, respectively. In the case of cysteine mutants, no enzyme activity was observed like tryptophan mutants, except for C139S. These results suggest that tryptophan and cysteine residues conserved in ST3Gal I are critical for its activity.
Description: 135-140
URI: http://hdl.handle.net/123456789/9809
ISSN: 0975-0959 (Online); 0301-1208 (Print)
Appears in Collections:IJBB Vol.47(3) [June 2010]

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