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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred


IMPACT FACTOR 2018: 3.014
5-year IMPACT FACTOR: 3.162

CiteScore 2018: 3.09

SCImago Journal Rank (SJR) 2018: 1.482
Source Normalized Impact per Paper (SNIP) 2018: 0.820

Online
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1437-4315
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Volume 390, Issue 3

Issues

Determination of three amino acids causing alteration of proteolytic activities of staphylococcal glutamyl endopeptidases

Takayuki K. Nemoto
  • 1Department of Oral Molecular Biology, Course of Medical and Dental Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588, Japan
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/ Toshio Ono
  • 2Department of Oral Molecular Biology, Course of Medical and Dental Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588, Japan
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/ Yu Shimoyama
  • 3Department of Oral Microbiology, Iwate Medical University School of Dentistry, Morioka 020-8505, Japan
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/ Shigenobu Kimura
  • 4Department of Oral Microbiology, Iwate Medical University School of Dentistry, Morioka 020-8505, Japan
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/ Yuko Ohara-Nemoto
  • 5Department of Oral Molecular Biology, Course of Medical and Dental Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588, Japan
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  • De Gruyter OnlineGoogle Scholar
Published Online: 2008-12-17 | DOI: https://doi.org/10.1515/BC.2009.027

Abstract

Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus warneri secrete glutamyl endopeptidases, designated GluV8, GluSE, and GluSW, respectively. The order of their protease activities is GluSE<GluSW<<GluV8. In the present study, we investigated the mechanism that causes these differences. Expression of chimeric proteins between GluV8 and GluSE revealed that the difference is primarily attributed to amino acid residues 170–195, which define the intrinsic protease activity, and additionally to residues 119–169, which affect the proteolytic sensitivity. Among nine substitutions present in residues 170–195 of the three proteases, the substitutions at positions 185, 188, and 189 were responsible for the changes in their activities, and the combination of W185, V188, and P189, which naturally occurs in GluV8, exerts the highest protease activity. W185 and P189 were indispensable for full activity, but V188 could be replaced by hydrophobic amino acids. These three amino acid residues appear to create a substrate-binding pocket together with the catalytic triad and the N-terminal V1, and therefore define the K m values of the proteases. We also describe a method to produce a chimeric form of GluSE and GluV8 that is resistant to proteolysis, and therefore possesses 4-fold higher activity than the wild-type recombinant GluV8.

Keywords: glutamyl endopeptidase; Staphylococcus aureus; Staphylococcus epidermidis; substrate-binding pocket; V8 protease

About the article

Corresponding author


Received: 2008-10-13

Accepted: 2008-11-29

Published Online: 2008-12-17

Published in Print: 2009-03-01


Citation Information: Biological Chemistry, Volume 390, Issue 3, Pages 277–285, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2009.027.

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[1]
Chen Chen, Vengadesan Krishnan, Kevin Macon, Kartik Manne, Sthanam V. L. Narayana, and Olaf Schneewind
Journal of Biological Chemistry, 2013, Volume 288, Number 41, Page 29440
[2]
Shakh M.A. Rouf, Yuko Ohara-Nemoto, Toshio Ono, Yu Shimoyama, Shigenobu Kimura, and Takayuki K. Nemoto
FEBS Open Bio, 2013, Volume 3, Number 1, Page 177
[3]
Toshio Ono, Yuko Ohara-Nemoto, Yu Shimoyama, Hisami Okawara, Takeshi Kobayakawa, Tomomi T. Baba, Shigenobu Kimura, and Takayuki K. Nemoto
Biological Chemistry, 2010, Volume 391, Number 10

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