PMID: 8604304Feb 15, 1996Paper

Mismatch DNA recognition protein from an extremely thermophilic bacterium, Thermus thermophilus HB8

Nucleic Acids Research
S TakamatsuSeiki Kuramitsu

Abstract

The mutS gene, implicated in DNA mismatch repair, was cloned from an extremely thermophilic bacterium, Thermus thermophilus HB8. Its nucleotide sequence encoded a 819-amino acid protein with a molecular mass of 91.4 kDa. Its predicted amino acid sequence showed 56 and 39% homology with Escherichia coli MutS and human hMsh2 proteins, respectively. The T.thermophilus mutS gene complemented the hypermutability of the E.coli mutS mutant, suggesting that T.thermophilus MutS protein was active in E.coli and could interact with E.coli MutL and/or MutH proteins. The T.thermophilus mutS gene product was overproduced in E.coli and then purified to homogeneity. Its molecular mass was estimated to be 91 kDa by SDS-PAGE but approx. 330 kDa by size-exclusion chromatography, suggesting that T.thermophilus MutS protein was a tetramer in its native state. Circular dichroic measurements indicated that this protein had an alpha-helical content of approx. 50%, and that it was stable between pH 1.5 and 12 at 25 degree C and was stable up to 80 degree C at neutral pH. Thermus thermophilus MutS protein hydrolyzed ATP to ADP and Pi, and its activity was maximal at 80 degrees C. The kinetic parameters of the ATPase activity at 65 degrees C were Km = 13...Continue Reading

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Citations

Mar 3, 2007·Analytical and Bioanalytical Chemistry·Michal MasaríkMiroslav Fojta
Jun 27, 2001·Mutation Research·P Hsieh
Mar 26, 2003·Protein Expression and Purification·Anna Stanisławska-SachadynJózef Kur
Jun 16, 2009·The Journal of Biological Chemistry·Lei TianGuo-Min Li
Oct 8, 2008·The Journal of Biological Chemistry·Kenji FukuiSeiki Kuramitsu
Dec 15, 1996·Nucleic Acids Research·N C NelsonM M Becker
Feb 26, 2005·Nucleic Acids Research·Melissa A CalmannM G Marinus
Aug 21, 2010·Journal of Nucleic Acids·Kenji Fukui
Oct 29, 2010·Journal of Nucleic Acids·Rihito MoritaSeiki Kuramitsu
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Aug 29, 2006·Journal of Biotechnology·Paweł SachadynJózef Kur
Sep 15, 2005·Journal of Biotechnology·Anna Stanisławska-SachadynJózef Kur
Mar 23, 2013·International Journal of Molecular Sciences·Kenji FukuiSeiki Kuramitsu
Jul 17, 2001·The Journal of Biological Chemistry·L J BlackwellP Modrich
Aug 24, 2017·The FEBS Journal·Koki OhshitaTaisuke Wakamatsu
Jun 28, 2003·The Journal of Biological Chemistry·Keith P BjornsonPaul Modrich
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Mar 4, 2008·The Journal of Biological Chemistry·Kenji FukuiSeiki Kuramitsu
Sep 5, 1997·The Journal of Biological Chemistry·N NakagawaS Kuramitsu
Aug 9, 2001·Journal of Molecular Biology·R KatoS Kuramitsu

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