title: PlyC: a multimeric bacteriophage lysin.
abstract: Lysins are murein hydrolases produced by bacteriophage that act on the bacterial host cell wall to release progeny phage. When added extrinsically in their purified form, these enzymes produce total lysis of susceptible Gram-positive bacteria within seconds, suggesting a unique antimicrobial strategy. All known Gram-positive lysins are produced as a single polypeptide containing a catalytic activity domain, which cleaves one of the four major peptidoglycan bonds, and a cell-wall-binding domain, which may bind a species-specific carbohydrate epitope in the cell wall. Here, we have cloned and expressed a unique lysin from the streptococcal bacteriophage C(1), termed PlyC. Molecular characterization of the plyC operon reveals that PlyC is, surprisingly, composed of two separate gene products, PlyCA and PlyCB. Based on biochemical and biophysical studies, the catalytically active PlyC holoenzyme is composed of eight PlyCB subunits for each PlyCA. Inhibitor studies predicted the presence of an active-site cysteine, and bioinformatic analysis revealed a cysteine, histidine-dependent amidohydrolase/peptidase domain within PlyCA. Point mutagenesis confirmed that PlyCA is responsible for the observed catalytic activity, and Cys-333 and His-420 are the active-site residues. PlyCB was found to self-assemble into an octamer, and this complex alone was able to direct streptococcal cell-wall-specific binding. Similar to no other proteins in sequence databases, PlyC defines a previously uncharacterized structural family of cell-wall hydrolases.
authors.affiliation: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. nelsond@rockefeller.edu
authors.forename: Daniel
authors.initials: D
authors.lastname: Nelson
authors.forename: Raymond
authors.initials: R
authors.lastname: Schuch
authors.forename: Peter
authors.initials: P
authors.lastname: Chahales
authors.forename: Shiwei
authors.initials: S
authors.lastname: Zhu
authors.forename: Vincent A
authors.initials: VA
authors.lastname: Fischetti
mesh_terms: D002473:Cell Wall / Q000201:enzymology / Q000235:genetics; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D017108:Streptococcus Phages / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics
chemical_list: D004798:Enzymes; C028806:lysin, streptococcus bacteriophage; D009238:N-Acetylmuramoyl-L-alanine Amidase