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Journal of Bacteriology, June 2000, p. 3383-3393, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Salmonella enterica Serovar Typhimurium Peptidase B Is a Leucyl Aminopeptidase with Specificity for Acidic Amino Acids

Zacharia Mathew, Tina M. Knox, and Charles G. Miller*

Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801

Received 10 January 2000/Accepted 18 March 2000

Peptidase B (PepB) of Salmonella enterica serovar Typhimurium is one of three broad-specificity aminopeptidases found in this organism. We have sequenced the pepB gene and found that it encodes a 427-amino-acid (46.36-kDa) protein, which can be unambiguously assigned to the leucyl aminopeptidase (LAP) structural family. PepB has been overexpressed and purified. The active enzyme shows many similarities to other members of the LAP family: it is a heat-stable (70°C; 20 min) hexameric (~270-kDa) metallopeptidase with a pH optimum of 8.5 to 9.5. A detailed study of the substrate specificity of the purified protein shows that it differs from other members of the family in its ability to hydrolyze peptides with N-terminal acidic residues. The preferred substrates for PepB are peptides with N-terminal Asp or Glu residues. Comparison of the amino acid sequence of PepB with those of other LAPs leads to the conclusion that PepB is the prototype of a new LAP subfamily with representatives in several other eubacterial species and to the prediction that the members of this family share the ability to hydrolyze peptides with N-terminal acidic residues. Site-directed mutagenesis has been used to show that this specificity appears to be determined by a single Lys residue present in a sequence motif conserved in all members of the subfamily.


* Corresponding author. Mailing address: Department of Microbiology, University of Illinois at Urbana-Champaign, B103 CLSL, 601 S. Goodwin Ave., Urbana, IL 61801. Phone: (217) 244-8418. Fax: (217) 244-6697. E-mail: charlesm{at}uiuc.edu.


Journal of Bacteriology, June 2000, p. 3383-3393, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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