PMID:20190047

Citation	Camiade, E, Peltier, J, Bourgeois, I, Couture-Tosi, E, Courtin, P, Antunes, A, Chapot-Chartier, MP, Dupuy, B and Pons, JL (2010) Characterization of Acp, a peptidoglycan hydrolase of Clostridium perfringens with N-acetylglucosaminidase activity that is implicated in cell separation and stress-induced autolysis. J. Bacteriol. 192:2373-84
Abstract	This work reports the characterization of the first known peptidoglycan hydrolase (Acp) produced mainly during vegetative growth of Clostridium perfringens. Acp has a modular structure with three domains: a signal peptide domain, an N-terminal domain with repeated sequences, and a C-terminal catalytic domain. The purified recombinant catalytic domain of Acp displayed lytic activity on the cell walls of several Gram-positive bacterial species. Its hydrolytic specificity was established by analyzing the Bacillus subtilis peptidoglycan digestion products by coupling reverse phase-high-pressure liquid chromatography (RP-HPLC) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, which displayed an N-acetylglucosaminidase activity. The study of acp expression showed a constant expression during growth, which suggested an important role of Acp in growth of C. perfringens. Furthermore, cell fractionation and indirect immunofluorescence staining using anti-Acp antibodies revealed that Acp is located at the septal peptidoglycan of vegetative cells during exponential growth phase, indicating a role in cell separation or division of C. perfringens. A knockout acp mutant strain was obtained by using the insertion of mobile group II intron strategy (ClosTron). The microscopic examination indicated a lack of vegetative cell separation in the acp mutant strain, as well as the wild-type strain incubated with anti-Acp antibodies, demonstrating the critical role of Acp in cell separation. The comparative responses of wild-type and acp mutant strains to stresses induced by Triton X-100, bile salts, and vancomycin revealed an implication of Acp in autolysis induced by these stresses. Overall, Acp appears as a major cell wall N-acetylglucosaminidase implicated in both vegetative growth and stress-induced autolysis.
Links	PubMed PMC2863477 Online version:10.1128/JB.01546-09
Keywords	Acetylglucosaminidase/genetics; Acetylglucosaminidase/metabolism; Acetylglucosaminidase/physiology; Anti-Bacterial Agents/pharmacology; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Bacterial Proteins/physiology; Bacteriolysis/drug effects; Bacteriolysis/genetics; Blotting, Western; Chromatography, High Pressure Liquid; Clostridium perfringens/cytology; Clostridium perfringens/drug effects; Clostridium perfringens/enzymology; Clostridium perfringens/genetics; Electrophoresis, Polyacrylamide Gel; Genome, Bacterial/genetics; Microbial Sensitivity Tests; Molecular Sequence Data; Mutagenesis; N-Acetylmuramoyl-L-alanine Amidase/genetics; N-Acetylmuramoyl-L-alanine Amidase/metabolism; N-Acetylmuramoyl-L-alanine Amidase/physiology; Octoxynol/pharmacology; Peptidoglycan/metabolism; Reverse Transcriptase Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
CLOPE:Q8XL11		GO:0061784: peptidoglycan N-acetylglucosaminidase activity	ECO:0000314:		F		Figure 4: Its hydrolytic specificity was established by analyzing the Bacillus subtilis peptidoglycan digestion products by coupling reverse phase-high-pressure liquid chromatography (RP-HPLC) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, which displayed an N-acetylglucosaminidase activity.	complete CACAO 12199

References

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