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Methods in Enzymology 1995
Topics: Amino Acid Sequence; Metalloendopeptidases; Molecular Sequence Data; Pseudomonas
PubMed: 7674963
DOI: 10.1016/0076-6879(95)48053-6 -
Methods in Enzymology 1995
Topics: Amino Acid Sequence; Escherichia coli; Insulin; Metalloendopeptidases; Molecular Sequence Data; Substrate Specificity
PubMed: 7674955
DOI: 10.1016/0076-6879(95)48045-5 -
Archives of Microbiology Apr 2006Metalloendopeptidase was isolated from Streptomyces rimosus culture filtrates in a homogeneous form. It was determined to be a 15 kDa basic protein, most active around...
Metalloendopeptidase was isolated from Streptomyces rimosus culture filtrates in a homogeneous form. It was determined to be a 15 kDa basic protein, most active around pH 7.5, and susceptible to inhibition by chelating agents, N-bromosuccinimide, thiorphan, and 10(-4) M zinc. The enzyme was highly specific for phenylalanine at the N-side of endopeptide bonds. Determination of amino acid sequence of the enzyme's NH(2)-part allowed the recognition of its structure homology with isolated and predicted metallopeptidases from several Streptomyces species. The data contribute to the definition of M7 family of metalloendopeptidases in streptomycetes.
Topics: Amino Acid Sequence; Culture Media; Hydrogen-Ion Concentration; Metalloendopeptidases; Molecular Sequence Data; Phenylalanine; Streptomyces; Substrate Specificity
PubMed: 16521041
DOI: 10.1007/s00203-006-0084-x -
Biochemistry. Biokhimiia Aug 2016Features of gene expression of the secreted Bacillus pumilus metalloendopeptidase belonging to the adamalysin/reprolysin family were investigated. In the regulatory...
Features of gene expression of the secreted Bacillus pumilus metalloendopeptidase belonging to the adamalysin/reprolysin family were investigated. In the regulatory region of the gene, we identified hypothetical binding sites for transcription factors CcpA and TnrA. We found that the expression of the metalloendopeptidase gene is controlled by mechanisms of carbon and nitrogen catabolite repression. In experiments involving nitrogen metabolism regulatory protein mutant strains, we found that the control of the metalloendopeptidase gene expression involves proteins of ammonium transport GlnK and AmtB interacting with the TnrA-regulator.
Topics: Bacillus pumilus; Bacterial Proteins; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Metalloendopeptidases; Nitrogen; Transcription Factors
PubMed: 27677556
DOI: 10.1134/S0006297916080095 -
Journal of Proteomics Jun 2020Snakebite envenoming affects millions of people worldwide, being officially considered a neglected tropical disease by the World Health Organization. The antivenom is...
Snakebite envenoming affects millions of people worldwide, being officially considered a neglected tropical disease by the World Health Organization. The antivenom is effective in neutralizing the systemic effects of envenomation, but local effects are poorly neutralized, often leading to permanent disability. The natural resistance of the South American pit viper Bothrops jararaca to its venom is partly attributed to BJ46a, a natural snake venom metalloendopeptidase inhibitor. Upon complex formation, BJ46a binds non-covalently to the metalloendopeptidase, rendering it unable to exert its proteolytic activity. However, the structural features that govern this interaction are largely unknown. In this work, we applied structural mass spectrometry techniques (cross-linking-MS and hydrogen-deuterium exchange MS) and in silico analyses (molecular modeling, docking, and dynamics simulations) to understand the interaction between BJ46a and jararhagin, a metalloendopeptidase from B. jararaca venom. We explored the distance restraints generated from XL-MS experiments to guide the modeling of BJ46a and jararhagin, as well as the protein-protein docking simulations. HDX-MS data pinpointed regions of protection/deprotection at the interface of the BJ46a-jararhagin complex which, in addition to the molecular dynamics simulation data, reinforced our proposed interaction model. Ultimately, the structural understanding of snake venom metalloendopeptidases inhibition by BJ46a could lead to the rational design of drugs to improve anti-snake venom therapeutics, alleviating the high morbidity rates currently observed.
Topics: Animals; Bothrops; Crotalid Venoms; Mass Spectrometry; Metalloendopeptidases; Bothrops jararaca Venom
PubMed: 32247172
DOI: 10.1016/j.jprot.2020.103761 -
Methods in Enzymology 1995
Topics: Amino Acid Sequence; Animals; Female; Humans; Matrix Metalloproteinase 7; Metalloendopeptidases; Molecular Sequence Data; Rats; Substrate Specificity; Uterus
PubMed: 7674940
DOI: 10.1016/0076-6879(95)48031-5 -
Methods in Enzymology 1995
Topics: Amino Acid Sequence; Crystallography, X-Ray; Kinetics; Metalloendopeptidases; Molecular Sequence Data; Substrate Specificity
PubMed: 7674929
DOI: 10.1016/0076-6879(95)48021-8 -
Rapid Communications in Mass... Jan 2020Lys-N, also known as lysine-specific metalloendopeptidase, functions as the "sister" enzyme of lysyl endopeptidase (Lys-C) in proteomic research. Its digestion...
RATIONALE
Lys-N, also known as lysine-specific metalloendopeptidase, functions as the "sister" enzyme of lysyl endopeptidase (Lys-C) in proteomic research. Its digestion specificity at the N-terminal lysine residue makes it a very useful tool in proteomics analysis, especially in mass spectrometry (MS)-based de novo sequencing of proteins.
METHODS
Here we present a complete production process of highly purified Lys-N from dry fruit of Grifola frondosa (maitake mushroom). The purification process includes one step of microfiltration plus one step of UF/DF (ultrafiltration used in tandem with a diafiltration method) recovery and four steps of chromatographic purification.
RESULTS
The overall yield of the process was approximately 6.7 mg Lys-N protein/kg dry fruit of G. frondosa. The assay data demonstrated that the purified Lys-N exhibited high enzymatic activity and specificity.
CONCLUSIONS
The novel production process provides for the first time the extraction of Lys-N from dry fruit of G. frondosa. The process is also stable and scalable, and provides an economic way of producing the enzyme in large quantities for MS-based proteomics and other biological studies.
Topics: Digestion; Fruiting Bodies, Fungal; Fungal Proteins; Grifola; Metalloendopeptidases; Proteomics; Serine Endopeptidases
PubMed: 31484223
DOI: 10.1002/rcm.8573 -
Annals of the New York Academy of... Jan 2013β-Lactam antibiotics are the most commonly used antibacterial agents and growing resistance to these drugs is a concern. Metallo-β-lactamases are a diverse set of... (Review)
Review
β-Lactam antibiotics are the most commonly used antibacterial agents and growing resistance to these drugs is a concern. Metallo-β-lactamases are a diverse set of enzymes that catalyze the hydrolysis of a broad range of β-lactam drugs including carbapenems. This diversity is reflected in the observation that the enzyme mechanisms differ based on whether one or two zincs are bound in the active site that, in turn, is dependent on the subclass of β-lactamase. The dissemination of the genes encoding these enzymes among Gram-negative bacteria has made them an important cause of resistance. In addition, there are currently no clinically available inhibitors to block metallo-β-lactamase action. This review summarizes the numerous studies that have yielded insights into the structure, function, and mechanism of action of these enzymes.
Topics: Catalysis; Catalytic Domain; Metalloendopeptidases; Mutagenesis; Protein Binding; Substrate Specificity; beta-Lactamases
PubMed: 23163348
DOI: 10.1111/j.1749-6632.2012.06796.x -
Neurochemical Research Nov 2018The mechanisms by which peptidergic signals are terminated have been the center of multiple studies leading to the discoveries of novel proteolytic activities. When... (Review)
Review
The mechanisms by which peptidergic signals are terminated have been the center of multiple studies leading to the discoveries of novel proteolytic activities. When studying the catabolic fate of neurotensin (NT) in brain and gastrointestinal tract, we detected a novel activity belonging to the metallopeptidases class and apparently distinct from previously known enzymes. Purification and cloning confirmed that this NT-degrading neutral metalloendopeptidase activity was indeed original. It was named endopeptidase 3.4.24.16 according to the IUBMB nomenclature and later, referred to as neurolysin. This review tells the history of neurolysin from its initial detection to its purification, cloning, design of specific inhibitors as well as in vitro and in vivo pharmacological studies aimed at delineating its role in the control of NT function. Finally, we discuss very recent advances suggesting a potential role of neurolysin in pathologies.
Topics: Amino Acid Sequence; Animals; Brain; Gastrointestinal Tract; Humans; Metalloendopeptidases; Neurotensin; Peptide Fragments
PubMed: 30159819
DOI: 10.1007/s11064-018-2624-6