PMID: 9171379Jun 1, 1997Paper

Identification of essential amino acids in the Streptococcus mutans glucosyltransferases

Journal of Bacteriology
H TsumoriH K Kuramitsu

Abstract

A comparison of the amino acid sequences of the glucosyltransferases (GTFs) of mutans streptococci with those from the alpha-amylase family of enzymes revealed a number of conserved amino acid positions which have been implicated as essential in catalysis. Utilizing a site-directed mutagenesis approach with the GTF-I enzyme of Streptococcus mutans GS-5, we identified three of these conserved amino acid positions, Asp413, Trp491, and His561, as being important in enzymatic activity. Mutagenesis of Asp413 to Thr resulted in a GTF which expressed only about 12% of the wild-type activity. In contrast, mutagenesis of Asp411 did not inhibit enzyme activity. In addition, the D413T mutant was less stable than was the parental enzyme when expressed in Escherichia coli. Moreover, conversion of Trp491 or His561 to either Gly or Ala resulted in enzymes devoid of GTF activity, indicating the essential nature of these two amino acids for activity. Furthermore, mutagenesis of the four Tyr residues present at positions 169 to 172 which are part of a subdomain with homology to the direct repeating sequences present in the glucan-binding domain of the GTFs had little overall effect on enzymatic activity, although the glucan products appeared to ...Continue Reading

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Citations

Mar 13, 1998·Journal of Dental Research·R A Burne
Jun 24, 2000·European Journal of Biochemistry·V MonchoisR R Russell
May 11, 1999·FEMS Microbiology Reviews·V MonchoisP Monsan
Jan 6, 2006·Archives of Microbiology·Pei-Min ChenJean-San Chia
Aug 23, 2002·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·D J Smith
Oct 21, 2000·FEMS Microbiology Letters·S JanecekR R Russell
Apr 16, 1999·Oral Microbiology and Immunology·G Hajishengallis, S M Michalek
Sep 25, 2004·Bioscience, Biotechnology, and Biochemistry·Kazumi FunaneMikihiko Kobayashi
Mar 10, 2006·Microbiology and Molecular Biology Reviews : MMBR·Sacha A F T van HijumIneke G H van Geel-Schutten
May 2, 2000·Applied and Environmental Microbiology·V MonchoisR R Russell
Oct 28, 2019·Biochemistry and Biophysics Reports·Shabeer A RatherAkhtar Mahmood
Mar 23, 2021·Biochemistry and Biophysics Reports·Dimple GoyalSukesh Chander Sharma
Mar 1, 2006·Biochimica Et Biophysica Acta·Wataru SaburiAtsuo Kimura
Aug 24, 2016·Journal of Agricultural and Food Chemistry·Xiao Yan Chen, Michael G Gänzle
May 19, 2009·Vaccine·Jinghua SunMingwen Fan

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