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

Biochem J. 1993 Jun 1;292 ( Pt 2)(Pt 2):563-70. doi: 10.1042/bj2920563.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacillus / enzymology*
  • Bacillus / physiology
  • Base Sequence
  • Carboxypeptidases / classification*
  • Carboxypeptidases / genetics
  • Carboxypeptidases / metabolism
  • Carboxypeptidases A
  • Cattle
  • Cloning, Molecular
  • DNA, Bacterial
  • Detergents
  • Endopeptidases / classification*
  • Endopeptidases / genetics
  • Endopeptidases / metabolism
  • Genes, Bacterial
  • Molecular Sequence Data
  • Peptide Mapping
  • Sequence Homology, Amino Acid
  • Spectrum Analysis
  • Spores, Bacterial
  • Zinc / metabolism*

Substances

  • DNA, Bacterial
  • Detergents
  • Carboxypeptidases
  • Endopeptidases
  • Carboxypeptidases A
  • gamma-D-glutamyl-meso-diaminopimelate peptidase I
  • Zinc

Associated data

  • GENBANK/D17642
  • GENBANK/D17799
  • GENBANK/D17800
  • GENBANK/D17801
  • GENBANK/D17802
  • GENBANK/D25299
  • GENBANK/D25301
  • GENBANK/D25302
  • GENBANK/X69507
  • GENBANK/X71774