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Proteolytic enzymes

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https://www.readbyqxmd.com/read/29157802/identification-and-in-silico-characterization-of-a-novel-peptide-inhibitor-of-angiotensin-converting-enzyme-from-pigeon-pea-cajanus-cajan
#1
K A Ayub Nawaz, Swapna Merlin David, Easwaran Murugesh, Murugesan Thandeeswaran, Kalarikkal Gopikrishnan Kiran, Ramasamy Mahendran, Muthusamy Palaniswamy, Jayaraman Angayarkanni
BACKGROUND: Plants are important sources of bioactive peptides. Among these, angiotensin converting enzyme (ACE) inhibitory peptides have a major focus on their ability to prevent hypertension. Inhibition of ACE has been established as an effective approach for the treatment of ACE associated diseases. HYPOTHESIS/PURPOSE: Some synthetic ACE inhibitory drugs cause side effects and hence there is a constant interest in natural compounds as alternatives. STUDY DESIGN: The study was designed to identify and characterize a peptide molecule from pigeon pea which has the biological property to inhibit ACE and can be developed as a therapeutic approach towards hypertension...
December 1, 2017: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/29155201/biochemical-properties-of-the-htra-homolog-from-bacterium-stenotrophomonas-maltophilia
#2
Urszula Zarzecka, Anna Modrak-Wojcik, Martyna Bayassi, Maciej Szewczyk, Artur Gieldon, Adam Lesner, Tomasz Koper, Agnieszka Bzowska, Maurizio Sanguinetti, Steffen Backert, Barbara Lipinska, Joanna Skorko-Glonek
The HtrA proteins due to their proteolytic, and in many cases chaperone activity, efficiently counteract consequences of stressful conditions. In the environmental bacterium and nosocomial pathogen Stenotrophomonas maltophilia HtrA (HtrASm) is induced as a part of adaptive response to host temperature (37°C). We examined the biochemical properties of HtrASm and compared them with those of model HtrAEc from Escherichia coli. We found that HtrASm is a protease and chaperone that operates over a wide range of pH and is highly active at temperatures between 35-37°C...
November 16, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29155158/thermodynamic-investigation-of-an-alkaline-protease-from-aspergillus-tamarii-urm4634-a-comparative-approach-between-crude-extract-and-purified-enzyme
#3
Osmar Soares da Silva, Rodrigo Lira de Oliveira, Jonatas de Carvalho Silva, Attilio Converti, Tatiana Souza Porto
The thermostable crude proteolytic extract and purified protease produced by Aspergillus tamarii URM4634 were investigated at different temperatures. The activity results were used to estimate the activation energy of the hydrolysis reaction catalyzed by crude extract and purified protease (E*=34.2 and 16.2kJ/mol) as well as the respective standard enthalpy variations of reversible enzyme unfolding (ΔH°u=31.9 and 13.9kJ/mol). When temperature was raised from 50 to 80°C in residual activity tests, the specific rate constant of crude proteolytic extract thermoinactivation increased from 0...
November 15, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29153522/short-communication-growth-of-dairy-isolates-of-geobacillus-thermoglucosidans-in-skim-milk-depends-on-lactose-degradation-products-supplied-by-anoxybacillus-flavithermus-as-secondary-species
#4
Y Zhao, M Kumar, M P M Caspers, M N Nierop Groot, J M B M van der Vossen, T Abee
Thermophilic bacilli such as Anoxybacillus and Geobacillus are important contaminants in dairy powder products. Remarkably, one of the common contaminants, Geobacillus thermoglucosidans, showed poor growth in skim milk, whereas significant growth of G. thermoglucosidans was observed in the presence of an Anoxybacillus flavithermus dairy isolate. In the present study, we investigated the underlying reason for this growth dependence of G. thermoglucosidans. Whole-genome sequences of 4 A. flavithermus strains and 4 G...
November 15, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/29153516/determination-of-lactadherin-concentration-in-dairy-by-products-by-elisa-effect-of-heat-treatment-and-hydrolysis
#5
Daniel Ripollés, José A Parrón, Javier Fraguas, Miguel Calvo, María D Pérez, Lourdes Sánchez
Lactadherin is a peripheral glycoprotein of the milk fat globule membrane with several attributed biological activities. In this study, we developed an indirect competitive ELISA to determine lactadherin concentration by using a rabbit polyclonal antiserum. The ELISA was applied to quantify lactadherin in several dairy by-products. Of the products tested, raw and commercial buttermilk had the highest concentrations of lactadherin (6.79 and 5.27 mg/g of product, respectively), followed by commercial butter serum (4...
November 15, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/29152044/structure-based-discovery-of-thiosemicarbazone-metalloproteinase-inhibitors-for-hemorrhage-treatment-in-snakebites
#6
Francis B Ferreira, Thiago M Pereira, Dayane L N Souza, Daiana S Lopes, Vitor Freitas, Veridiana M R Ávila, Arthur E Kümmerle, Carlos Mauricio R Sant'Anna
The venoms of snakes are composed by many toxins, which are responsible for various toxic effects including intense pain, bleeding disorders, and local tissue damage caused by hemorrhage and necrosis. The snake venom metalloproteinases (SVMPs) are proteolytic zinc-dependent enzymes acting in different hemostatic mechanisms. In this work, a structure-based molecular modeling strategy was used for the rational design, by means of a homology 3D model of an SVMP isolated from Bothrops pauloensis venom (BpMP-I), followed by synthesis and in vitro evaluation of new thiosemicarbazones as the first inhibitors of the B...
November 9, 2017: ACS Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/29150754/detoxification-of-aflatoxin-b1-by-antifungal-compounds-from-lactobacillus-brevis-and-lactobacillus-paracasei-isolated-from-dairy-products
#7
Eman Zakaria Gomaa, Manal Farouk Abdelall, Omima Mohammed El-Mahdy
Aflatoxins are a large group of highly toxic, mutagenic, and carcinogenic mycotoxins produced by specific species of fungi. Potential contamination of food commodities by these compounds causes extensive damage that lead to great economic losses. This study explored the potential use of antifungal compounds, produced by Lactobacillus brevis and Lactobacillus paracasei, for growth inhibition and subsequent aflatoxin B1 production from select strains of Aspergillus flavus and Aspergillus parasiticus. Lactobacilli strains were isolated from traditional Egyptian dairy products, whereas fungal strains were isolated from infected cereal seeds...
November 18, 2017: Probiotics and Antimicrobial Proteins
https://www.readbyqxmd.com/read/29147551/development-of-proteolytically-stable-n-methylated-peptide-inhibitors-of-aggregation-of-the-amylin-peptide-implicated-in-type-2-diabetes
#8
Idira Obasse, Mark Taylor, Nigel J Fullwood, David Allsop
Islet amyloid polypeptide, also known as amylin, is the main component of the amyloid deposits present in approximately 90% of people with type 2 diabetes mellitus (T2DM). In this disease, amylin aggregates into multimeric β-pleated sheet structures which cause damage to pancreatic islet β-cells. Inhibitors of early-stage amylin aggregation could therefore provide a disease-modifying treatment for T2DM. In this study, overlapping peptides were designed to target the 'binding' region (RLANFLVHSS, residues 11-20) of human amylin, and their effects on amyloid fibril formation were determined by thioflavin-T assay...
December 6, 2017: Interface Focus
https://www.readbyqxmd.com/read/29145643/the-merops-database-of-proteolytic-enzymes-their-substrates-and-inhibitors-in-2017-and-a-comparison-with-peptidases-in-the-panther-database
#9
Neil D Rawlings, Alan J Barrett, Paul D Thomas, Xiaosong Huang, Alex Bateman, Robert D Finn
The MEROPS database (http://www.ebi.ac.uk/merops/) is an integrated source of information about peptidases, their substrates and inhibitors. The hierarchical classification is: protein-species, family, clan, with an identifier at each level. The MEROPS website moved to the EMBL-EBI in 2017, requiring refactoring of the code-base and services provided. The interface to sequence searching has changed and the MEROPS protein sequence libraries can be searched at the EMBL-EBI with HMMER, FastA and BLASTP. Cross-references have been established between MEROPS and the PANTHER database at both the family and protein-species level, which will help to improve curation and coverage between the resources...
November 14, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29144877/role-of-proteolytic-enzyme-inhibitors-on-carious-and-eroded-dentin-associated-with-a-universal-bonding-system
#10
M C Giacomini, Pmc Scaffa, L P Chaves, Cmp Vidal, T N Machado, H M Honório, L Tjäderhane, L Wang
OBJECTIVES: The aim of this study was to evaluate the effect of proteolytic inhibitors on the bond strength of a universal adhesive system (etch-and-rinse mode) applied to artificial carious and eroded dentin. METHODS: Ninety molars were prepared and randomly divided into three groups according to the substrate: N, no challenges; ACD, artificial carious dentin simulation and ERO, artificial erosion simulation with orange juice. All groups were redivided into three subgroups according to the dentin pretreatment: W, water; CHX, 2% digluconate chlorhexidine; and E-64 (trans-epoxysuccinyl-L-leucylamido-[4-guanidino] butane), 5 μM E-64 inhibitor...
November 2017: Operative Dentistry
https://www.readbyqxmd.com/read/29144294/higher-alkyl-sulfatase-activity-required-by-microbial-inhabitants-to-remove-anionic-surfactants-in-the-contaminated-surface-waters
#11
Bulent Icgen, Salih Batuhan Salik, Lale Goksu, Huseyin Ulusoy, Fadime Yilmaz
Biodegradation of anionic surfactants, like sodium dodecyl sulfate (SDS) are challenged by some bacteria through the function of the enzyme alkyl sulfatases. Therefore, identifying and characterizing bacteria capable of degrading SDS with high alkyl sulfatase enzyme activity are pivotal. In this study, bacteria isolated from surfactant contaminated river water were screened for their potential to degrade SDS. Primary screening carried out by the conventional enrichment culture technique and assessment of SDS-degrading ability through methylene blue active substance assay revealed 12, out of 290, SDS-degrading surface water bacteria with maximum SDS degrading abilities of 46-94% in 24-54 h...
November 2017: Water Science and Technology: a Journal of the International Association on Water Pollution Research
https://www.readbyqxmd.com/read/29142432/inhibition-of-glycation-induced-cytotoxicity-protein-glycation-and-activity-of-proteolytic-enzymes-by-extract-from-perovskia-atriplicifolia-roots
#12
Mehran Miroliaei, Akram Aminjafari, Sylwester Ślusarczyk, Izabela Nawrot-Hadzik, Mehdi Rahimmalek, Adam Matkowski
Background: Protein glycation and glycotoxicity belong to the main oxidative-stress related complications in diabetes. Perovskia species are used in Asian folk medicine as antidiabetic herbs. Objective: The aim of this study was to verify the ability of the methanolic extract from Perovskia atriplicifolia Benth. roots to diminish glycation of albumin and to prevent cell damage in vitro. Furthermore, we tested the extract for in vitro antioxidant activity and inhibition of elastase and collagenase...
October 2017: Pharmacognosy Magazine
https://www.readbyqxmd.com/read/29139623/extracellular-proteinases-of-candida-species-pathogenic-yeasts
#13
Maria Rapala-Kozik, Oliwia Bochenska, Dorota Zajac, Justyna Karkowska-Kuleta, Mariusz Gogol, Marcin Zawrotniak, Andrzej Kozik
The increased incidence of severe disseminated infections caused by opportunistic yeast-like fungi Candida spp. highlights the urgent need for research into the major virulence factors of these pathogens - extracellular aspartic proteinases of the candidapepsin and yapsin families. Classically, these enzymes were considered to be generally destructive factors that damage host tissues and provide nutrients for pathogen propagation. However, in recent decades, novel and more specific functions have been suggested for extracellular candidal proteinases...
November 15, 2017: Molecular Oral Microbiology
https://www.readbyqxmd.com/read/29132916/insulin-degrading-enzyme-in-the-fight-against-alzheimer-s-disease
#14
REVIEW
Igor V Kurochkin, Enrico Guarnera, Igor N Berezovsky
After decades of research and clinical trials there is still no cure for Alzheimer's disease (AD). While impaired clearance of amyloid beta (Aβ) peptides is considered as one of the major causes of AD, it was recently complemented by a potential role of other toxic amyloidogenic species. Insulin-degrading enzyme (IDE) is the proteolytic culprit of various β-forming peptides, both extracellular and intracellular. On the basis of demonstrated allosteric activation of IDE against Aβ, it is possible to propose a new strategy for the targeted IDE-based cleansing of different toxic aggregation-prone peptides...
November 10, 2017: Trends in Pharmacological Sciences
https://www.readbyqxmd.com/read/29132905/purification-and-biochemical-characterization-of-a-22-kda-stable-cysteine-like-protease-from-the-excretory-secretory-product-of-the-liver-fluke-fasciola-hepatica-by-using-conventional-techniques
#15
Ahmed Hemici, Roumaila Sabrina Benerbaiha, Dalila Bendjeddou
This study describes the purification and characterization of a stable protease activity isolated from Fasciola hepatica adult worms maintained in vitro by employing acetone precipitation (40-60%) followed by a gel filtration through Sephadex G-100 and DEAE- cellulose ion exchange column. Through this three-step purification, the enzyme was purified 11-fold with a specific activity of 1893.9U/mg and 31.5% recovery. After the final ultrafiltration step, the purification fold was increased up to 13.1 and the overall activity yield reached a rate of 18...
October 28, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29129910/thermostable-exoshells-fold-and-stabilize-recombinant-proteins
#16
Siddharth Deshpande, Nihar D Masurkar, Vallerinteavide Mavelli Girish, Malan Desai, Goutam Chakraborty, Juliana M Chan, Chester L Drum
The expression and stabilization of recombinant proteins is fundamental to basic and applied biology. Here we have engineered a thermostable protein nanoparticle (tES) to improve both expression and stabilization of recombinant proteins using this technology. tES provides steric accommodation and charge complementation to green fluorescent protein (GFPuv), horseradish peroxidase (HRPc), and Renilla luciferase (rLuc), improving the yields of functional in vitro folding by ~100-fold. Encapsulated enzymes retain the ability to metabolize small-molecule substrates, presumably via four 4...
November 13, 2017: Nature Communications
https://www.readbyqxmd.com/read/29129076/comprehensive-molecular-characterization-of-a-cisplatin-specific-monoclonal-antibody
#17
Lena Ruhe, Stefanie Ickert, Ulrike Hochkirch, Johanna Hofmann, Sebastian Beck, Jürgen Thomale, Michael W Linscheid
Despite their immense and rapidly increasing importance as analytical tools or therapeutic drugs, the detailed structural features of particular monoclonal antibodies are widely unknown. Here, an antibody already in use for diagnostic purposes and for molecular dosimetry studies in cancer therapy with very high affinity and specificity for cisplatin-induced DNA modifications was studied extensively. The molecular structure and modifications as well as the antigen specificity were investigated mainly by mass spectrometry...
November 11, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/29128226/camel-milk-protein-hydrolysates-with-improved-technofunctional-properties-and-enhanced-antioxidant-potential-in-in-vitro-and-in-food-model-systems
#18
Kholoud Awad Al-Shamsi, Priti Mudgil, Hassan Mohamed Hassan, Sajid Maqsood
Camel milk protein hydrolysates (CMPH) were generated using proteolytic enzymes, such as alcalase, bromelain, and papain, to explore the effect on the technofunctional properties and antioxidant potential under in vitro and in real food model systems. Characterization of the CMPH via degree of hydrolysis, sodium dodecyl sulfate-PAGE, and HPLC revealed that different proteins in camel milk underwent degradation at different degrees after enzymatic hydrolysis using 3 different enzymes for 2, 4, and 6 h, with papain displaying the highest degradation...
November 8, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/29126465/facile-synthesis-of-magnetic-metal-organic-frameworks-for-highly-efficient-proteolytic-digestion-used-in-mass-spectrometry-based-proteomics
#19
Rui Zhai, Yifeng Yuan, Fenglong Jiao, Feiran Hao, Xiang Fang, Yangjun Zhang, Xiaohong Qian
Mass spectrometry (MS) based bottom-up strategy is now the first choice for proteomics analysis. In this process, highly efficient and complete enzymatic degradation of protein samples is extremely important to achieve in-depth protein coverage and high-throughput protein profiling. However, conventional in-solution digestion suffer from long digestion time and enzyme autolysis that limit the protein sample processing throughput and identification accuracy. Here, we developed a novel type of magnetic metal organic frameworks (MOFs)-based immobilized enzyme reactor, Fe3O4@DOTA-ZIF-90-trypsin...
November 22, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/29122614/a-recombinant-fusion-protein-based-fluorescent-protease-assay-for-high-throughput-compatible-substrate-screening
#20
Beáta Bozóki, Lívia Gazda, Ferenc Tóth, Márió Miczi, János András Mótyán, József Tőzsér
In connection with the intensive investigation of proteases, several methods have been developed for analysis of the substrate specificity. Due to the great number of proteases and the expected target molecules to be analyzed, time- and cost-efficient high-throughput screening (HTS) methods are preferred. Here we describe the development and application of a separation-based HTS-compatible fluorescent protease assay, which is based on the use of recombinant fusion proteins as substrates of proteases. The protein substrates used in this assay consists of N-terminal (hexahistidine and maltose binding protein) fusion tags, cleavage sequences of the tobacco etch virus (TEV) and HIV-1 proteases, and a C-terminal fluorescent protein (mApple or mTurquoise2)...
November 6, 2017: Analytical Biochemistry
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