Structure and activity of ChiX: a peptidoglycan hydrolase required for chitinase secretion by
Autor: | Richard A, Owen, Paul K, Fyfe, Adam, Lodge, Jacob, Biboy, Waldemar, Vollmer, William N, Hunter, Frank, Sargent |
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Rok vydání: | 2017 |
Předmět: |
Models
Molecular Protein Conformation alpha-Helical anomalous dispersion crystal structure peptidoglycan hydrolase Amino Acid Motifs Genetic Vectors Gene Expression Chitin Peptidoglycan Crystallography X-Ray Substrate Specificity Bacterial Proteins Operon Escherichia coli Protein Interaction Domains and Motifs Cloning Molecular Serratia marcescens Research Articles bacterial chitinase secretion Aspartic Acid Binding Sites zinc enzyme Hydrolysis Chitinases Gene Expression Regulation Bacterial N-Acetylmuramoyl-L-alanine Amidase Recombinant Proteins Zinc Protein Conformation beta-Strand mutagenesis Protein Binding Research Article |
Zdroj: | Biochemical Journal |
ISSN: | 1470-8728 |
Popis: | The Gram-negative bacterium Serratia marcescens secretes many proteins that are involved in extracellular chitin degradation. This so-called chitinolytic machinery includes three types of chitinase enzymes and a lytic polysaccharide monooxygenase. An operon has been identified in S. marcescens, chiWXYZ, that is thought to be involved in the secretion of the chitinolytic machinery. Genetic evidence points to the ChiX protein being a key player in the secretion mechanism, since deletion of the chiX gene in S. marcescens led to a mutant strain blocked for secretion of all members of the chitinolytic machinery. In this work, a detailed structural and biochemical characterisation of ChiX is presented. The high-resolution crystal structure of ChiX reveals the protein to be a member of the LAS family of peptidases. ChiX is shown to be a zinc-containing metalloenzyme, and in vitro assays demonstrate that ChiX is an l-Ala d-Glu endopeptidase that cleaves the cross-links in bacterial peptidoglycan. This catalytic activity is shown to be intimately linked with the secretion of the chitinolytic machinery, since substitution of the ChiX Asp-120 residue results in a variant protein that is both unable to digest peptidoglycan and cannot rescue the phenoytype of a chiX mutant strain. |
Databáze: | OpenAIRE |
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