Characterization of the sporulation-related gamma-D-glutamyl-(L)meso-diaminopimelic-acid-hydrolysing peptidase I of Bacillus sphaericus NCTC 9602 as a member of the metallo(zinc) carboxypeptidase A family. Modular design of the protein.
The sporulation-related gamma-D-glutamyl-(L)meso-diaminopimelic-acid-hydrolysing peptidase I of Bacillus sphaericus NCTC 9602 has been analysed by proton-induced X-ray emission. It contains 1 equivalent Zn2+ per mol of protein. As derived from gene cloning and sequencing, the B. sphaericus Zn peptidase I is a two-module protein. A 100-amino-acid-residue N-terminal domain consisting of two tandem segments of similar sequences, is fused to a 296-amino-acid-residue C-terminal catalytic domain. The catalytic domain belongs to the Zn carboxypeptidase A family, the closest match being observed with the Streptomyces griseus carboxypeptidase [Narahashi (1990) J. Biochem. 107, 879-886] and with the family prototype, bovine carboxypeptidase A. The catalytic domain of the B. sphaericus peptidase I possesses, distributed along the amino-acid sequence, peptide segments, a triad His162-Glu165-His307 and a dyad Tyr347-Glu366 that are equivalent to secondary structures, the zinc-binding triad His69-Glu72-His196 and the catalytic dyad Tyr248-Glu270 of bovine carboxypeptidase A respectively. The N-terminal repeats of the B. sphaericus peptidase I have similarity with the C-terminal repeats of the Enterococcus hirae muramidase 2, the Streptococcus (now Enterococcus) faecalis autolysin and the Bacillus phi PZA and phi 29 lysozymes, to which a role in the recognition of a particular moiety of the bacterial cell envelope has been tentatively assigned. Detergents enhance considerably the specific activity of the B. sphaericus peptidase I.
Hourdou
M L
ML
Centre d'Ingénierie des Protéines, Université de Liège, Belgium.
Guinand
M
M
Vacheron
M J
MJ
Michel
G
G
Denoroy
L
L
Duez
C
C
Englebert
S
S
Joris
B
B
Weber
G
G
Ghuysen
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000201:enzymology* / Q000502:physiology; D001483:Base Sequence; D002268:Carboxypeptidases / Q000145:classification* / Q000235:genetics / Q000378:metabolism; D043422:Carboxypeptidases A; D002417:Cattle; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D003902:Detergents; D010450:Endopeptidases / Q000145:classification* / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D017386:Sequence Homology, Amino Acid; D013057:Spectrum Analysis; D013171:Spores, Bacterial; D015032:Zinc / Q000378:metabolism*
D004269:DNA, Bacterial; D003902:Detergents; D002268:Carboxypeptidases; D010450:Endopeptidases; D043422:Carboxypeptidases A; C019404:gamma-D-glutamyl-meso-diaminopimelate peptidase I; D015032:Zinc