RRC ID 15640
著者 Yokoi KJ, Sugahara K, Iguchi A, Nishitani G, Ikeda M, Shimada T, Inagaki N, Yamakawa A, Taketo A, Kodaira K.
タイトル Molecular properties of the putative autolysin Atl(WM) encoded by Staphylococcus warneri M: mutational and biochemical analyses of the amidase and glucosaminidase domains.
ジャーナル Gene
Abstract The putative autolysin Atl(WM) of Staphylococcus warneri M is a modular protein exhibiting two enzyme activities, an N-terminal side amidase (ami(atlwm)-R1-R2) and a C-terminal side glucosaminidase (R3-glu(atlwm)). Zymographic analysis of the protein overproduced in Escherichia coli showed that both enzymes were active toward 17 Gram-positive bacteria, including staphylococci, lactobacilli, lactococci, enterococci, and micrococci. The purified enzyme core ami(atlwm) (or glu(atlwm)) had the pH and temperature optima of about 7.0 (5.5) and 41 (50) degrees C, respectively. ami(atlwm) was inactivated by EDTA, and was stimulated by such salts as CoCl(2), MnCl(2), CaCl(2), or ZnCl(2). Six mutations within ami(atlwm), (H362A, E421A, H467A, H479, D481A, and Y491D) drastically reduced cell-lytic activity. Comparative analysis with other related amidases suggested that the three residues H362, H467, and D481 likely act as ligands (and/or active sites). The lytic activity of glu(atlwm) markedly declined in four mutants (E1238A, E1238Q, T1239A, and Y1332A). For determination of the putative cell-recognition regions, four domains (R1-R2, R1, R2, and R3) were purified; all the proteins substantially bound to S. warneri M cells from exponential to stationary growth phases, and R1-R2 aggregated the cells. Protein sequencing and immunoblot analysis suggested that the extacellular Atl(WM) might be primarily processed at two specific sites (one between pro and ami(atlwm), and the other between R2 and R3) to yield the mature amidase and glucosaminidase.
巻・号 416(1-2)
ページ 66-76
公開日 2008-6-15
DOI 10.1016/j.gene.2008.03.004
PII S0378-1119(08)00115-7
PMID 18440165
MeSH Amidohydrolases / analysis* Amidohydrolases / genetics* Amino Acid Sequence Bacterial Proteins / analysis Bacterial Proteins / genetics DNA Mutational Analysis Escherichia coli / genetics Gram-Positive Bacteria / drug effects Hexosaminidases / analysis* Hexosaminidases / genetics* Hydrogen-Ion Concentration Molecular Sequence Data N-Acetylmuramoyl-L-alanine Amidase / analysis* N-Acetylmuramoyl-L-alanine Amidase / genetics* Staphylococcus / genetics* Temperature Transfection
IF 2.984
引用数 26
WOS 分野 GENETICS & HEREDITY
リソース情報
一般微生物 JCM 1130 JCM 2151 JCM 2416 JCM 2418 JCM 2423 JCM 2424