Identification of endo-beta-N-acetylglucosaminidase and N-acetylmuramyl-L-alanine amidase as cluster-dispersing enzymes in Staphylococcus aureus

Sugai, Motoyuki; Komatsuzawa, Hitoshi; Akiyama, Tomoko; Hong, Yeong‐Man; Oshida, Tadahiro; Miyake, Yoichiro; Yamaguchi, Takayuki; Suginaka, Hidekazu

doi:10.1128/jb.177.6.1491-1496.1995

Cited by 117 publications

(132 citation statements)

References 30 publications

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“…SDS-polyacrylamide gel electrophoresis and Western blotting were carried out as described elsewhere (26). Immunodetection of protein was performed with the ECL Western blotting analysis system (Amersham Pharmacia).…”

Section: Methodsmentioning

confidence: 99%

“…The IPG gel was placed on top of the stacking gel, and ran for 7 hr at 30 mA. The electrophoresis and staining with Coomassie brilliant blue were performed as already described (26).…”

Section: Methodsmentioning

confidence: 99%

“…The 51 kDa GL and the 62 kDa AM were shown to disperse clusters of S. aureus without affecting cell viability (26). The atl gene products including 51 kDa GL and 62 kDa AM were found to form a ring structure on the cell surface at the septal region for the upcoming cell division site (30).…”

mentioning

confidence: 99%

“…These mutants grew normally and did not form giant clusters when grown in broth culture (6,26), but only formed small clusters. This mutant phenotype contradicted with the notion that atl is involved in cell separation because one would expect the mutant to form giant clusters if the daughter cells are unable to separate.…”

mentioning

confidence: 99%

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Molecular Characterization of an atl Null Mutant of Staphylococcus aureus

Takahashi

Komatsuzawa

Yamada

et al. 2002

Microbiology and Immunology

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atl is a gene encoding a bifunctional peptidoglycan hydrolase of Staphylococcus aureus. The gene product of atl is a 138 kDa protein that has an amidase domain and a glucosaminidase domain, and undergoes processing to generate two major peptidoglycan hydrolases, a 51 kDa glucosaminidase and a 62 kDa amidase in culture supernatant. An atl null mutant was isolated by allelic replacement and characterized. The mutant grew in clusters and sedimented when grown in broth culture. Analysis of peptidoglycan prepared from the wild type and the mutant revealed that there were no differences in muropeptide composition or in glycan chain length distribution. On the other hand, the atl mutation resulted in pleiotropic effects on cell surface nature. The mutant cells showed complete inhibition of metabolic turnover of cell wall peptidoglycan and revealed a rough outer cell wall surface. The mutation also decreased the amount of protein non‐covalently bound to the cell surface and altered the protein profile, but did not affect proteins covalently associated with the cell wall. Lysis of growing cells treated with otherwise lytic concentration of penicillin G was completely inhibited in the mutant, but that of non‐growing cells was not affected by the mutation. The atl mutation did not significantly affect the ability of S. aureus to provoke an acute infection when inoculated intraperitoneally in a mouse sepsis model. These results further support the supposition that atl gene products are involved in cell separation, cell wall turnover and penicillin‐induced lysis of the cells.

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Section: Methodsmentioning

confidence: 99%

“…The IPG gel was placed on top of the stacking gel, and ran for 7 hr at 30 mA. The electrophoresis and staining with Coomassie brilliant blue were performed as already described (26).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Molecular Characterization of an atl Null Mutant of Staphylococcus aureus

Takahashi

Komatsuzawa

Yamada

et al. 2002

Microbiology and Immunology

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“…Purified LukF and LukS were provided by Dr. M. G. Bowden (Texas A&M University System Health Science Center, Houston, TX) after purification by ion-exchange chromatography followed by liquid chromatography (22). Antiserum from rabbits against amidase (60 kDa), glucosaminidase (50 kDa), the complete autolysin molecule (138 kDa, amidase and glucosamidase), and as control anti-epidermal differentiation factors A were used as previously described (23). Antiserum from rabbits against Panton-Valentine leukocidin (PVL)-F, PVL-S, PVL-FS, and as control plasmin sensitive IgG1 and clumping factor A IgG 1004 were used as previously described (24).…”

Section: Specific S Aureus Reagentsmentioning

confidence: 99%

A Novel Mechanism of Rapid Nuclear Neutrophil Extracellular Trap Formation in Response to Staphylococcus aureus

Pilsczek

Salina

Poon

et al. 2010

The Journal of Immunology

924

945

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Neutrophil extracellular traps (NETs) are webs of DNA covered with antimicrobial molecules that constitute a newly described killing mechanism in innate immune defense. Previous publications reported that NETs take up to 3–4 h to form via an oxidant-dependent event that requires lytic death of neutrophils. In this study, we describe neutrophils responding uniquely to Staphylococcus aureus via a novel process of NET formation that did not require neutrophil lysis or even breach of the plasma membrane. The multilobular nucleus rapidly became rounded and condensed. During this process, we observed the separation of the inner and outer nuclear membranes and budding of vesicles, and the separated membranes and vesicles were filled with nuclear DNA. The vesicles were extruded intact into the extracellular space where they ruptured, and the chromatin was released. This entire process occurred via a unique, very rapid (5–60 min), oxidant-independent mechanism. Mitochondrial DNA constituted very little if any of these NETs. They did have a limited amount of proteolytic activity and were able to kill S. aureus. With time, the nuclear envelope ruptured, and DNA filled the cytoplasm presumably for later lytic NET production, but this was distinct from the vesicular release mechanism. Panton–Valentine leukocidin, autolysin, and a lipase were identified in supernatants with NET-inducing activity, but Panton–Valentine leukocidin was the dominant NET inducer. We describe a new mechanism of NET release that is very rapid and contributes to trapping and killing of S. aureus.

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Antibacterial Property of Oxygen‐Terminated Carbon Bonds

Yang

Zhang

Wang

et al. 2022

Adv Funct Materials

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Inspired by the antibacterial agents, such as ethylene oxide, anacardic acid, and salicylic acid, oxygen‐terminated carbon bonds may take the major responsibility in antibacterial properties, a layer of oxygen‐terminated carbon bonds is fabricated on nanodiamond as an immobilized coating for the investigation of the antibacterial activity. The results demonstrate comparable antibacterial activities on oxygen‐terminated nanodiamond coating (NDC) against two kinds of probe bacteria, Gram‐negative Escherichia coli and Gram‐positive Staphylococcus aureus, and the annealing method as a proper method for the improvement of the antibacterial performance of the NDC. The concentration of oxygen‐terminated carbon bonds, especially CO and CO, and the ratio of CO to CO, are found to determine the antibacterial performance of NDCs. The antibacterial behavior performed through the surface interaction is revealed and the abnormal expression of cell wall‐related proteins are identified, where a potential antibacterial mechanism is proposed: the oxygen‐terminated carbon bonds damaged the cell wall through the blockage and the electrostatic repulsive force, which aggravated the abnormal metabolism of bacteria and lead to its death. The antibacterial property of oxygen‐terminated carbon bonds is confirmed and its application, NDC, provides a new idea for antibacterial research and opens the door to antibacterial coatings for medical devices.

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Identification of endo-beta-N-acetylglucosaminidase and N-acetylmuramyl-L-alanine amidase as cluster-dispersing enzymes in Staphylococcus aureus

Cited by 117 publications

References 30 publications

Molecular Characterization of an atl Null Mutant of Staphylococcus aureus

Molecular Characterization of an atl Null Mutant of Staphylococcus aureus

A Novel Mechanism of Rapid Nuclear Neutrophil Extracellular Trap Formation in Response to Staphylococcus aureus

Antibacterial Property of Oxygen‐Terminated Carbon Bonds

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