Multiple Enzymatic Activities of the Murein Hydrolase from Staphylococcal Phage φ11

Navarre, William Wiley; Ton‐That, Hung; Faull, Kym F.; Schneewind, Olaf

doi:10.1074/jbc.274.22.15847

Cited by 167 publications

(178 citation statements)

References 62 publications

(78 reference statements)

Supporting

Mentioning

173

Contrasting

Order By: Relevance

“…All previously reported lysins derived from Gram-positiveinfecting bacteriophage are composed of one or more (22) catalytic domains linked to a binding domain and are synthesized as single polypeptides. Thus, PlyC is an unusual example of a multimeric lysin composed of two distinct gene products, PlyCA and PlyCB.…”

Section: Discussionmentioning

confidence: 99%

PlyC: A multimeric bacteriophage lysin

Nelson

Schuch

Chahales

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

190

189

View full text Add to dashboard Cite

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 Grampositive 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 C1, 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.multimeric protein ͉ cell-wall hydrolase ͉ Streptococcus

show abstract

Section: Discussionmentioning

confidence: 99%

PlyC: A multimeric bacteriophage lysin

Nelson

Schuch

Chahales

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

190

189

View full text Add to dashboard Cite

show abstract

“…However, when studied at the biochemical level, most of these "extra" domains were found to be silent. For example, the staphylococcal Φ11 lysin has both D-alanyl-glycyl endopeptidase and N-acetylmuramoyl-Lalanine amidase catalytic domains (6). Although the endopeptidase domain was active by itself (20,21), the amidase domain was shown to be silent by deletion analysis (21).…”

Section: Discussionmentioning

confidence: 99%

“…In some rare cases, two to three catalytic domains may be linked to a single binding domain (6,7). With few exceptions, the catalytic domain is usually represented by one of four families of peptidoglycan hydrolases: N-acetylglucosaminidases, N-acetylmuramidases (lysozymes), N-acetylmuramoyl-L-alanine amidases, and endopeptidases (8).…”

mentioning

confidence: 99%

X-ray crystal structure of the streptococcal specific phage lysin PlyC

McGowan

Buckle

Mitchell

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

134

View full text Add to dashboard Cite

Bacteriophages deploy lysins that degrade the bacterial cell wall and facilitate virus egress from the host. When applied exogenously, these enzymes destroy susceptible microbes and, accordingly, have potential as therapeutic agents. The most potent lysin identified to date is PlyC, an enzyme assembled from two components (PlyCA and PlyCB) that is specific for streptococcal species. Here the structure of the PlyC holoenzyme reveals that a single PlyCA moiety is tethered to a ring-shaped assembly of eight PlyCB molecules. Structure-guided mutagenesis reveals that the bacterial cell wall binding is achieved through a cleft on PlyCB. Unexpectedly, our structural data reveal that PlyCA contains a glycoside hydrolase domain in addition to the previously recognized cysteine, histidine-dependent amidohydrolases/peptidases catalytic domain. The presence of eight cell wall-binding domains together with two catalytic domains may explain the extraordinary potency of the PlyC holoenyzme toward target bacteria.

show abstract

“…19,24,25 For instance, the staphylococcal phage phi11 endolysin has both an endopeptidase and an amidase. 26 Likewise, the streptococcal phage endolysin B30 has both an endopeptidase and a muramidase EAD. 27 However, sitedirected mutagenesis and deletion analysis of dual EAD enzymes has indicated that one of the EAD tends to be dominant and highly catalytic, whereas the other is virtually inactive in vitro.…”

Section: Endolysins -Peptidoglycan Degrading Enzymesmentioning

confidence: 99%