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Research Article

Biochemical Characterization of the First Essential Two-Component Signal Transduction System from Staphylococcus aureus and Streptococcus pneumoniae

Clausen V.A. · Bae W. · Throup J. · Burnham M.K.R. · Rosenberg M. · Wallis N.G.

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Antimicrobials and Host Defense, GlaxoSmithKline Pharmaceuticals, Collegeville, Pa., USA

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J Mol Microbiol Biotechnol 2003;5:252–260

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Article / Publication Details

First-Page Preview
Abstract of Research Article

Received: November 22, 2002
Accepted: March 21, 2003
Published online: July 16, 2003
Issue release date: July 2003

Number of Print Pages: 9
Number of Figures: 5
Number of Tables: 1

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: https://www.karger.com/MMB

Abstract

The yycFG two-component signal transduction system (TCSTS) has been shown to be essential to the viability of several gram-positive bacteria. However, the function of the gene pair remains unknown. Interestingly, while both components are essential to Staphylococcus aureus and Bacillus subtilis, only the response regulator (yycF) is essential to Streptococcus pneumoniae. To study this essential TCSTS further, the S. pneumoniae and S. aureus truncated YycG histidine kinase and full-length YycF response regulator proteins were characterized at a biochemical level. The recombinant proteins from both organisms were expressed in Escherichia coli and purified. The YycG autophosphorylation activities were activated by ammonium. The apparent Km (ATP) of S. aureus YycG autophosphorylation was 130 µM and S. pneumoniae was 3.0 µM. Each had similar kcat values of 0.036 and 0.024 min–1, respectively. Cognate phosphotransfer was also investigated indicating different levels of the phosphorylated YycG intermediates during the reaction. The S. pneumoniae YycG phosphorylated intermediate was not detectable in the presence of its cognate YycF, while phosphorylated S. aureus YycG and YycF were detected concurrently. In addition, noncognate phosphotransfer was demonstrated between the two species. These studies thoroughly compare the essential YycFG TCSTS from the two species at the biochemical level and also establish methods for assaying the activities of these antibacterial targets.

© 2003 S. Karger AG, Basel


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Article / Publication Details

First-Page Preview
Abstract of Research Article

Received: November 22, 2002
Accepted: March 21, 2003
Published online: July 16, 2003
Issue release date: July 2003

Number of Print Pages: 9
Number of Figures: 5
Number of Tables: 1

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: https://www.karger.com/MMB


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