The structure of the N-terminal module of the cell wall hydrolase RipA and its role in regulating catalytic activity

2018 ◽  
Vol 86 (9) ◽  
pp. 912-923 ◽  
Author(s):  
Eva Maria Steiner ◽  
Jeppe Lyngsø ◽  
Jodie E. Guy ◽  
Gleb Bourenkov ◽  
Ylva Lindqvist ◽  
...  
Keyword(s):  
Cell Wall ◽  
Catalytic Activity ◽  
Cell Wall Hydrolase

scholarly journals Identification and Characterization of a Peptidoglycan Hydrolase, MurA, of Listeria monocytogenes, a Muramidase Needed for Cell Separation

2003 ◽  
Vol 185 (23) ◽  
pp. 6801-6808 ◽  
Author(s):  
Shannon A. Carroll ◽  
Torsten Hain ◽  
Ulrike Technow ◽  
Ayub Darji ◽  
Philippos Pashalidis ◽  
...  
Keyword(s):  
Cell Wall ◽  
Listeria Monocytogenes ◽  
Cell Separation ◽  
Bacterial Species ◽  
Surface Protein ◽  
Wild Type ◽  
Terminal Domain ◽  
Cell Wall Hydrolase ◽  
Characteristic Features ◽  
Type Gene

ABSTRACT A novel cell wall hydrolase encoded by the murA gene of Listeria monocytogenes is reported here. Mature MurA is a 66-kDa cell surface protein that is recognized by the well-characterized L. monocytogenes-specific monoclonal antibody EM-7G1. MurA displays two characteristic features: (i) an N-terminal domain with homology to muramidases from several gram-positive bacterial species and (ii) four copies of a cell wall-anchoring LysM repeat motif present within its C-terminal domain. Purified recombinant MurA produced in Escherichia coli was confirmed to be an authentic cell wall hydrolase with lytic properties toward cell wall preparations of Micrococcus lysodeikticus. An isogenic mutant with a deletion of murA that lacked the 66-kDa cell wall hydrolase grew as long chains during exponential growth. Complementation of the mutant strain by chromosomal reintegration of the wild-type gene restored expression of this murein hydrolase activity and cell separation levels to those of the wild-type strain. Studies reported herein suggest that the MurA protein is involved in generalized autolysis of L. monocytogenes.

scholarly journals Antimicrobial Activity of Exebacase (Lysin CF-301) against the Most Common Causes of Infective Endocarditis

2019 ◽  
Vol 63 (10) ◽  
Author(s):  
Aubrey Watson ◽  
Jun Taek Oh ◽  
Karen Sauve ◽  
Patricia A. Bradford ◽  
Cara Cassino ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Cell Wall ◽  
Infective Endocarditis ◽  
Broth Microdilution ◽  
Coagulase Negative Staphylococci ◽  
Content Type ◽  
Microdilution Method ◽  
Cell Wall Hydrolase ◽  
Broth Microdilution Method ◽  
Common Causes

ABSTRACT Exebacase, a recombinantly produced lysin (cell wall hydrolase), and comparator antibiotics were tested by the broth microdilution method against strain sets of Staphylococcus and Streptococcus spp., which are the most common causes of infective endocarditis in humans. Exebacase was active against all Staphylococcus spp. tested, including S. aureus and coagulase-negative staphylococci (MIC50/90, 0.5/1 μg/ml). Activity against Streptococcus spp. was variable, with S. pyogenes, S. agalactiae, and S. dysgalactiae (MIC50/90, 1/2 μg/ml) among the most susceptible.

scholarly journals Functional analysis of Clostridium difficile sortase B reveals key residues for catalytic activity and substrate specificity

2020 ◽  
Vol 295 (11) ◽  
pp. 3734-3745
Author(s):  
Chia-Yu Kang ◽  
I-Hsiu Huang ◽  
Chi-Chi Chou ◽  
Tsai-Yu Wu ◽  
Jyun-Cyuan Chang ◽  
...  
Keyword(s):  
Cell Wall ◽  
Catalytic Activity ◽  
Clostridium Difficile ◽  
Substrate Specificity ◽  
Surface Proteins ◽  
Site Directed Mutagenesis ◽  
Serine Residue ◽  
Peptidoglycan Cell Wall ◽  
Attractive Therapeutic Target ◽  
Key Residues

Most of Gram-positive bacteria anchor surface proteins to the peptidoglycan cell wall by sortase, a cysteine transpeptidase that targets proteins displaying a cell wall sorting signal. Unlike other bacteria, Clostridium difficile, the major human pathogen responsible for antibiotic-associated diarrhea, has only a single functional sortase (SrtB). Sortase's vital importance in bacterial virulence has been long recognized, and C. difficile sortase B (Cd-SrtB) has become an attractive therapeutic target for managing C. difficile infection. A better understanding of the molecular activity of Cd-SrtB may help spur the development of effective agents against C. difficile infection. In this study, using site-directed mutagenesis, biochemical and biophysical tools, LC-MS/MS, and crystallographic analyses, we identified key residues essential for Cd-SrtB catalysis and substrate recognition. To the best of our knowledge, we report the first evidence that a conserved serine residue near the active site participates in the catalytic activity of Cd-SrtB and also SrtB from Staphylococcus aureus. The serine residue indispensable for SrtB activity may be involved in stabilizing a thioacyl-enzyme intermediate because it is neither a nucleophilic residue nor a substrate-interacting residue, based on the LC-MS/MS data and available structural models of SrtB–substrate complexes. Furthermore, we also demonstrated that residues 163–168 located on the β6/β7 loop of Cd-SrtB dominate specific recognition of the peptide substrate PPKTG. The results of this work reveal key residues with roles in catalysis and substrate specificity of Cd-SrtB.

scholarly journals Nucleotide sequence and regulation of a new putative cell wall hydrolase gene, cwlD, which affects germination in Bacillus subtilis. .

1995 ◽  
Vol 177 (19) ◽  
pp. 5582-5589 ◽  
Author(s):  
J Sekiguchi ◽  
K Akeo ◽  
H Yamamoto ◽  
F K Khasanov ◽  
J C Alonso ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Nucleotide Sequence ◽  
Cell Wall Hydrolase
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