The nonribosomal peptide synthetase HMWP2 forms a thiazoline ring during biogenesis of yersiniabactin, an iron-chelating virulence factor of Yersinia pestis.

Abstract

Pathogenic Yersinia species have been shown to synthesize a siderophore molecule, yersiniabactin, as a virulence factor during iron starvation. Here we provide the first biochemical evidence for the role of the Yersinia pestis high molecular weight protein 2 (HMWP2), a nonribosomal peptide synthetase homologue, and YbtE in the initiation of yersiniabactin biosynthesis. YbtE catalyzes the adenylation of salicylate and the transfer of this activated salicyl group to the N-terminal aryl carrier protein domain (ArCP; residues 1-100) of HMWP2. A fragment of HMWP2, residues 1-1491, can adenylate cysteine and with the resulting cysteinyl-AMP autoaminoacylate the peptidyl carrier protein domain (PCP1; residues 1383-1491) either in cis or in trans. Catalytic release of hydroxyphenylthiazoline carboxylic acid (HPT-COOH) and/or N-(hydroxyphenylthiazolinylcarbonyl)cysteine (HPT-cys) is observed upon incubation of YbtE, HMWP2 1-1491, L-cysteine, salicylate, and ATP. These products presumably arise from nucleophilic attack by water or cysteine of a stoichiometric hydroxyphenylthiazolinylcarbonyl-S-PCP1-HMWP2 intermediate. Detection of the heterocyclization capacity of HMWP2 1-1491 implies salicyl-transferring and thiazoline-forming activity for the HMWP2 condensation domain (residues 101-544) and is the first demonstration of such heterocyclization ability in a nonribosomal peptide synthetase enzyme.

• Authors: A M Gehring, I Mori, R D Perry, C T Walsh
• Journal: Biochemistry (Biochemistry) Vol. 37 Issue 33 Pg. 11637-50 (Aug 18 1998) ISSN: 0006-2960 [Print] United States
• PMID: 9709002 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Bacterial Outer Membrane Proteins
• Bacterial Proteins
• Carrier Proteins
• Iron-Binding Proteins
• Peptide Fragments
• Periplasmic Binding Proteins
• Phenols
• Siderophores
• Thiazoles
• yersiniabactin
• Adenosine Monophosphate
• Iron
• Peptide Synthases
• Cysteine

Topics

• Adenosine Monophosphate (metabolism)
• Bacterial Outer Membrane Proteins
• Bacterial Proteins (chemistry, metabolism)
• Carrier Proteins (metabolism)
• Catalysis
• Cysteine (metabolism)
• Iron (metabolism)
• Iron-Binding Proteins
• Peptide Fragments (chemical synthesis, metabolism)
• Peptide Synthases (chemistry, metabolism)
• Periplasmic Binding Proteins
• Phenols
• Protein Structure, Tertiary
• Siderophores (metabolism)
• Thiazoles (metabolism)
• Virulence
• Yersinia pestis (enzymology, pathogenicity)