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Eva Maria Steiner, Jeppe Lyngsø, Jodie E Guy, Gleb Bourenkov, Ylva Lindqvist, Thomas R Schneider, Jan Skov Pedersen, Gunter Schneider, Robert Schnell
RipA plays a vital role during cell division of Mycobacterium tuberculosis by degrading the cell wall peptidoglycan at the septum, allowing daughter cell separation. The peptidoglycan degrading activity relies on the NlpC/P60 domain, and as it is potentially harmful when deregulated, spatial and temporal control is necessary in this process. The N-terminal domain of RipA has been proposed to play an inhibitory role blocking the C-terminal NlpC/P60 domain. Accessibility of the active site cysteine residue is however not limited by the presence of the N-terminal domain, but by the lid-module of the inter-domain linker, which is situated in the peptide binding groove of the crystal structures of the catalytic domain...
May 2, 2018: Proteins
Tatsuya Fukushima, Natsuki Uchida, Masatoshi Ide, Takeko Kodama, Junichi Sekiguchi
Biopolymers on the cell surface are very important for protecting microorganisms from environmental stresses, as well as storing nutrients and minerals. Synthesis of biopolymers is well studied, while studies on the modification and degradation processes of biopolymers are limited. One of these biopolymers, poly-γ-glutamic acid (γ-PGA), is produced by Bacillus species. Bacillus subtilis PgdS, possessing three NlpC/P60 domains, hydrolyses γ-PGA. Here, we have demonstrated that several dl-endopeptidases with an NlpC/P60 domain (LytE, LytF, CwlS, CwlO, and CwlT) in B...
March 2018: Microbiology
Cleber Eduardo Galvão, Stenio Perdigão Fragoso, Carina Elisei de Oliveira, Odinéia Forner, Renata Ribeiro Bastos Pereira, Cleber Oliveira Soares, Grácia Maria Soares Rosinha
BACKGROUND: Caseous lymphadenitis (CLA) is a disease that affects sheep, goats and occasionally humans. The etiologic agent is the Corynebacterium pseudotuberculosis bacillus. The objective of this study was to build a gene expression library from C. pseudotuberculosis and use immunoscreening to identify genes that encode potential antigenic proteins for the development of DNA and subunit vaccines against CLA. RESULTS: A wild strain of C. pseudotuberculosis was used for extraction and partial digestion of genomic DNA...
September 21, 2017: BMC Microbiology
Vivian H Tang, Geoffrey A Stewart, Barbara J Chang
The bodies and faecal pellets of the house dust mite (HDM), Dermatophagoides pteronyssinus, are the source of many allergenic and nonallergenic proteins. One of these, the 14-kDa bacteriolytic enzyme LytFM, originally isolated from the spent HDM growth medium, may contribute to bacteriolytic activity previously detected by zymography at 14 kDa in the culture supernatants of some bacterial species isolated from surface-sterilised HDM. Based on previously reported findings of lateral gene transfer between microbes and their eukaryotic hosts, we investigated the presence of lytFM in the genomes of nine Gram-positive bacteria from surface-sterilised HDM, and the expression by these isolates of LytFM and its variants LytFM1/LytFM2...
September 2017: FEBS Open Bio
Se Kye Kim, Yun Min Park, Kyoung Hwa Jung, Young Gyu Chai
Bacillus anthracis, an aetiologic agent of the zoonotic disease anthrax, encodes a putative NlpC/P60 endopeptidase BAS1812. It harbours a signal peptide, three bacterial SH3 domains and an NlpC/P60 family domain. Previous studies showed that BAS1812 is immunogenic in infected hosts and is a potential biomarker for anthrax treatment. To date, however, little information is known about its function and involvement in anthrax pathogenesis. Here we describe the phenotypic effect of BAS1812 deletion in B. anthracis Sterne strain...
February 2017: Microbiology
Kavita J Rangan, Virginia A Pedicord, Yen-Chih Wang, Byungchul Kim, Yun Lu, Shai Shaham, Daniel Mucida, Howard C Hang
The intestinal microbiome modulates host susceptibility to enteric pathogens, but the specific protective factors and mechanisms of individual bacterial species are not fully characterized. We show that secreted antigen A (SagA) from Enterococcus faecium is sufficient to protect Caenorhabditis elegans against Salmonella pathogenesis by promoting pathogen tolerance. The NlpC/p60 peptidoglycan hydrolase activity of SagA is required and generates muramyl-peptide fragments that are sufficient to protect C. elegans against Salmonella pathogenesis in a tol-1-dependent manner...
September 23, 2016: Science
Minfeng Yu, Jing Yang, Minliang Guo
Lysin motif (LysM) is a highly conserved carbohydrate binding module that is widely present in proteins from both prokaryotes and eukaryotes. LysM domains from many LysM-containing proteins can be taken out of their natural context and retain their ability to bind peptidoglycan. Therefore, LysM has enormous potential for applications in both industry and medicine. This potential has stimulated an intensive search for LysM modules with different evolutionary origins. The p60 protein (Lm-p60) is an NlpC/P60-containing peptidoglycan hydrolase secreted by Listeria monocytogenes...
November 2016: Bioengineered
Avery E Sears, Krzysztof Palczewski
Lecithin:retinol acyltransferase (LRAT) catalyzes the acyl transfer from the sn-1 position of phosphatidylcholine (PC) to all-trans-retinol, creating fatty acid retinyl esters (palmitoyl, stearoyl, and some unsaturated derivatives). In the eye, these retinyl esters are substrates for the 65 kDa retinoid isomerase (RPE65). LRAT is well characterized biochemically, and recent structural data from closely related family members of the NlpC/P60 superfamily and a chimeric protein have established its catalytic mechanism...
June 7, 2016: Biochemistry
John P Bannantine, Cari K Lingle, Philip R Adam, Kasra X Ramyar, William J McWhorter, Judith R Stabel, William D Picking, Brian V Geisbrecht
A subset of proteins containing NlpC/P60 domains are bacterial peptidoglycan hydrolases that cleave noncanonical peptide linkages and contribute to cell wall remodeling as well as cell separation during late stages of division. Some of these proteins have been shown to cleave peptidoglycan in Mycobacterium tuberculosis and play a role in Mycobacterium marinum virulence of zebra fish; however, there are still significant knowledge gaps concerning the molecular function of these proteins in Mycobacterium avium subspecies paratuberculosis (MAP)...
April 2016: Protein Science: a Publication of the Protein Society
Mohammad Tahir Yousafzai, Naveed Zafar Janjua, Amna Rehana Siddiqui, Shafquat Rozi
AIM: We assessed the compliance at first level care facilities (FLCF) with universal precautions (UP) and its behavioral predictors using Health Belief Model (HBM). METHODS: A sample of FLCF from public clinic (PC), privately owned licensed practitioners' clinic (LPC) and non-licensed practitioners' clinic (NLPC) was obtained. Health Care Workers (HCW) who diagnose and prescribe medication was termed as Prescriber and that carries out prescriber's order was defined Assistant...
October 2015: International Journal of Health Sciences
Qingping Xu, Dominique Mengin-Lecreulx, Xueqian W Liu, Delphine Patin, Carol L Farr, Joanna C Grant, Hsiu-Ju Chiu, Lukasz Jaroszewski, Mark W Knuth, Adam Godzik, Scott A Lesley, Marc-André Elsliger, Ashley M Deacon, Ian A Wilson
UNLABELLED: Bacterial SH3 (SH3b) domains are commonly fused with papain-like Nlp/P60 cell wall hydrolase domains. To understand how the modular architecture of SH3b and NlpC/P60 affects the activity of the catalytic domain, three putative NlpC/P60 cell wall hydrolases were biochemically and structurally characterized. These enzymes all have γ-d-Glu-A2pm (A2pm is diaminopimelic acid) cysteine amidase (or dl-endopeptidase) activities but with different substrate specificities. One enzyme is a cell wall lysin that cleaves peptidoglycan (PG), while the other two are cell wall recycling enzymes that only cleave stem peptides with an N-terminal l-Ala...
September 15, 2015: MBio
A M Vyunishev, V G Arkhipkin, V V Slabko, I S Baturin, A R Akhmatkhanov, V Ya Shur, A S Chirkin
We study second-harmonic generation (SHG) of femtosecond laser pulses in a rectangular two-dimensional nonlinear photonic crystal (NLPC). Multiple SH beams were observed in the vicinity of the propagation direction of the fundamental beam. It has been verified that the angular positions of these beams obey the conditions of nonlinear Raman-Nath diffraction (NRND). The measured SH spectra of specific NRND orders consist of narrow peaks that experience a high-frequency spectral shift as the order grows. We derive an analytical expression for the process studied and find the theoretical results to be in good agreement with the experimental data...
September 1, 2015: Optics Letters
Catarina G Fernandes, Diana Plácido, Diana Lousa, José A Brito, Anabela Isidro, Cláudio M Soares, Jan Pohl, Maria A Carrondo, Margarida Archer, Adriano O Henriques
Transglutaminases are best known for their ability to catalyze protein cross-linking reactions that impart chemical and physical resilience to cellular structures. Here, we report the crystal structure and characterization of Tgl, a transglutaminase from the bacterium Bacillus subtilis. Tgl is produced during sporulation and cross-links the surface of the highly resilient spore. Tgl-like proteins are found only in spore-forming bacteria of the Bacillus and Clostridia classes, indicating an ancient origin. Tgl is a single-domain protein, produced in active form, and the smallest transglutaminase characterized to date...
September 22, 2015: Biochemistry
Jaslyn E M M Wong, Søren Roi Midtgaard, Kira Gysel, Mikkel B Thygesen, Kasper K Sørensen, Knud J Jensen, Jens Stougaard, Søren Thirup, Mickaël Blaise
LysM domains, which are frequently present as repetitive entities in both bacterial and plant proteins, are known to interact with carbohydrates containing N-acetylglucosamine (GlcNAc) moieties, such as chitin and peptidoglycan. In bacteria, the functional significance of the involvement of multiple LysM domains in substrate binding has so far lacked support from high-resolution structures of ligand-bound complexes. Here, a structural study of the Thermus thermophilus NlpC/P60 endopeptidase containing two LysM domains is presented...
March 2015: Acta Crystallographica. Section D, Biological Crystallography
Siqing Liu, Joseph O Rich, Amber Anderson
The cell-free supernatant (CFS) from Lactobacillus paracasei NRRL B-50314 culture has been previously reported as containing antibacterial activity against a wide variety of Gram-positive bacteria. The CFS protein gel slice corresponding to antibacterial activities was subjected to trypsin digestion and ion trap MASS (Gel/LC-MS/MS) analysis. BlastP search of the resulted IQAVISIAEQQIGKP sequence led to a hypothetical cell-wall associated hydrolase (designated as CWH here) from Lactobacillus paracasei ATCC 25302...
February 2015: Journal of Industrial Microbiology & Biotechnology
Qingping Xu, Dominique Mengin-Lecreulx, Delphine Patin, Joanna C Grant, Hsiu-Ju Chiu, Lukasz Jaroszewski, Mark W Knuth, Adam Godzik, Scott A Lesley, Marc-André Elsliger, Ashley M Deacon, Ian A Wilson
GlcNAc-1,6-anhydro-MurNAc-tetrapeptide is a major peptidoglycan degradation intermediate and a cytotoxin. It is generated by lytic transglycosylases and further degraded and recycled by various enzymes. We have identified and characterized a highly specific N-acetylmuramoyl-L-alanine amidase (AmiA) from Bacteroides uniformis, a member of the DUF1460 protein family, that hydrolyzes GlcNAc-1,6-anhydro-MurNAc-peptide into disaccharide and stem peptide. The high-resolution apo structure at 1.15 Å resolution shows that AmiA is related to NlpC/P60 γ-D-Glu-meso-diaminopimelic acid amidases and shares a common catalytic core and cysteine peptidase-like active site...
December 2, 2014: Structure
Jaslyn E M M Wong, Husam M A B Alsarraf, Jørn Døvling Kaspersen, Jan Skov Pedersen, Jens Stougaard, Søren Thirup, Mickaël Blaise
Cellulose, chitin and peptidoglycan are major long-chain carbohydrates in living organisms, and constitute a substantial fraction of the biomass. Characterization of the biochemical basis of dynamic changes and degradation of these β,1-4-linked carbohydrates is therefore important for both functional studies of biological polymers and biotechnology. Here, we investigated the functional role of multiplicity of the carbohydrate-binding lysin motif (LysM) domain that is found in proteins involved in bacterial peptidoglycan synthesis and remodelling...
February 2014: FEBS Journal
Ruth Y Eberhardt, Yuanyuan Chang, Alex Bateman, Alexey G Murzin, Herbert L Axelrod, William C Hwang, L Aravind
BACKGROUND: The NTF2-like superfamily is a versatile group of protein domains sharing a common fold. The sequences of these domains are very diverse and they share no common sequence motif. These domains serve a range of different functions within the proteins in which they are found, including both catalytic and non-catalytic versions. Clues to the function of protein domains belonging to such a diverse superfamily can be gleaned from analysis of the proteins and organisms in which they are found...
2013: BMC Bioinformatics
Jun Kurushima, Ikue Hayashi, Motoyuki Sugai, Haruyoshi Tomita
Enterococcus faecalis strains are commensal bacteria in humans and other animals, and they are also the causative agent of opportunistic infectious diseases. Bacteriocin 41 (Bac41) is produced by certain E. faecalis clinical isolates, and it is active against other E. faecalis strains. Our genetic analyses demonstrated that the extracellular products of the bacL1 and bacA genes, which are encoded in the Bac41 operon, coordinately express the bacteriocin activity against E. faecalis. In this study, we investigated the molecular functions of the BacL1 and BacA proteins...
December 27, 2013: Journal of Biological Chemistry
Jaslyn E M M Wong, Mickael Blaise
Autolysins belong to a protein family involved in peptidoglycan degradation and remodelling. Within this family, NlpC/P60 endopeptidases are involved in the hydrolysis of the peptide arm of peptidoglycan. In this work, the putative NlpC/P60 endopeptidase TTHA0266 from Thermus thermophilus HB8 was overexpressed, purified and crystallized. The crystals diffracted to 2.4 Å resolution and belonged to the hexagonal space group P6(1), with unit-cell parameters a = b = 71.19, c = 198.68 Å, γ = 120°. Selenomethionine-substituted protein was crystallized and the structure was solved by single-wavelength anomalous dispersion...
November 2013: Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
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