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https://www.readbyqxmd.com/read/28231319/development-of-a-keratinase-activity-assay-using-recombinant-chicken-feather-keratin-substrates
#1
Hyeon-Su Jin, Seon Yeong Park, Kyungmin Kim, Yong-Jik Lee, Gae-Won Nam, Nam Joo Kang, Dong-Woo Lee
Poultry feathers consist mainly of the protein keratin, which is rich in β-pleated sheets and consequently resistant to proteolysis. Although many keratinases have been identified, the reasons for their substrate specificity towards β-keratin remain unclear due to difficulties in preparing a soluble feather keratin substrate for use in activity assays. In the present study, we overexpressed Gallus gallus chromosomes 2 and 27 β-keratin-encoding genes in Escherichia coli, purified denatured recombinant proteins by Ni2+ affinity chromatography, and refolded by stepwise dialysis to yield soluble keratins...
2017: PloS One
https://www.readbyqxmd.com/read/28231306/effects-of-adamts14-genetic-polymorphism-and-cigarette-smoking-on-the-clinicopathologic-development-of-hepatocellular-carcinoma
#2
Ming-Jen Sheu, Ming-Ju Hsieh, Ying-Erh Chou, Po-Hui Wang, Chao-Bin Yeh, Shun-Fa Yang, Hsiang-Lin Lee, Yu-Fan Liu
BACKGROUND: ADAMTS14 is a member of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs), which are proteolytic enzymes with a variety of further ancillary domain in the C-terminal region for substrate specificity and enzyme localization via extracellular matrix association. However, whether ADAMTS14 genetic variants play a role in hepatocellular carcinoma (HCC) susceptibility remains unknown. METHODOLOGY/PRINCIPAL FINDINGS: Four non-synonymous single-nucleotide polymorphisms (nsSNPs) of the ADAMTS14 gene were examined from 680 controls and 340 patients with HCC...
2017: PloS One
https://www.readbyqxmd.com/read/28224230/newly-isolated-bacillus-sp-g51-from-patagonian-wool-produces-an-enzyme-combination-suitable-for-felt-resist-treatments-of-organic-wool
#3
Martín S Iglesias, Cynthia Sequeiros, Sebastián García, Nelda L Olivera
Bacteria from Patagonian Merino wool were isolated to assess their wool-keratinolytic activity and potential for felt-resist treatments. Strains from Bacillus, Exiguobacterium, Deinococcus, and Micrococcus produced wool-degrading enzymes. Bacillus sp. G51 showed the highest wool-keratinolytic activity. LC-MS/MS analysis revealed that G51 secreted two serine proteases belonging to the peptidase family S8 (MEROPS) and a metalloprotease associated with Bacillolysin, along with other enzymes (γ-glutamyltranspeptidase and dihydrolipoyl dehydrogenases) that could be involved in reduction of keratin disulfide bonds...
February 21, 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28223360/a-novel-subtilase-inhibitor-in-plants-shows-structural-and-functional-similarities-to-protease-propeptides
#4
Mathias Hohl, Annick Stintzi, Andreas Schaller
The propeptides of subtilisin-like serine proteinases (SBTs) serve dual functions as intramolecular chaperones that are required for enzyme folding and as inhibitors of the mature proteases. SBT propeptides are homologous to the I9 family of protease inhibitors that have only been described in fungi. Here we report the identification and characterization of Subtilisin Propeptide-Like Inhibitor 1 (SPI-1) from Arabidopsis thaliana. Sequence similarity and the shared β-α-β-β-α-β core structure identified SPI-1 as a member of the I9 inhibitor family, and as the first independent I9 inhibitor in higher eukaryotes...
February 21, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28223215/bufadienolides-from-kalanchoe-daigremontiana-as-thrombin-inhibitors-in-vitro-and-in-silico-study
#5
Joanna Kolodziejczyk-Czepas, Malgorzata Sieradzka, Barbara Moniuszko-Szajwaj, Łukasz Pecio, Michal B Ponczek, Pawel Nowak, Anna Stochmal
Thrombin is an active plasma coagulation factor II, critical for the formation of fibrin clot during blood coagulation. For that reason, this protein is also a crucial target for different anti-thrombotic therapies. The work is based on in vitro evaluation of the inhibitory effect of bufadienolide-rich fraction, isolated from roots of Kalanchoe daigremontiana (1-50μg/ml) on enzymatic properties of a serine proteinase - thrombin. The efficacy of the inhibition of amidolytic activity of thrombin (measured as a hydrolysis of the chromogenic substrate S-2238, Chromogenix) attained about 10 and 66%, respectively...
February 18, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28219449/ex-vivo-18-o-labeling-mass-spectrometry-identifies-a-peripheral-amyloid-%C3%AE-clearance-pathway
#6
Erik Portelius, Niklas Mattsson, Josef Pannee, Henrik Zetterberg, Magnus Gisslén, Hugo Vanderstichele, Eleni Gkanatsiou, Gabriela A N Crespi, Michael W Parker, Luke A Miles, Johan Gobom, Kaj Blennow
BACKGROUND: Proteolytic degradation of amyloid β (Aβ) peptides has been intensely studied due to the central role of Aβ in Alzheimer's disease (AD) pathogenesis. While several enzymes have been shown to degrade Aβ peptides, the main pathway of Aβ degradation in vivo is unknown. Cerebrospinal fluid (CSF) Aβ42 is reduced in AD, reflecting aggregation and deposition in the brain, but low CSF Aβ42 is, for unknown reasons, also found in some inflammatory brain disorders such as bacterial meningitis...
February 20, 2017: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/28217561/characterisation-of-the-thermostable-protease-aprx-in-strains-of-pseudomonas-fluorescens-and-impact-on-the-shelf-life-of-dairy-products-preliminary-results
#7
Nadia Andrea Andreani, Lisa Carraro, Luca Fasolato, Stefania Balzan, Rosaria Lucchini, Enrico Novelli, Barbara Cardazzo
Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial proteases, a predominant role in spoilage of dairy products seems to be played by the thermostable metallo-protease AprX, which is produced by various strains of Pseudomonas fluorescens. Differences in AprX enzyme activity among different strains were highlighted, but the most proteolytic strains were not identified. In this study, the presence of the aprX gene was evaluated in 69 strains isolated from food matrices and 18 reference strains belonging to the P...
September 20, 2016: Italian Journal of Food Safety
https://www.readbyqxmd.com/read/28217537/effect-of-proteolytic-enzyme-bromelain-on-pain-and-swelling-after-removal-of-third-molars
#8
Tejpal Singh, Vinayak More, Umayra Fatima, Tanveer Karpe, Mohammed A Aleem, J Prameela
AIMS AND OBJECTIVES: To evaluate the effect of bromelain on pain and swelling after surgical removal of third molars. MATERIALS AND METHODS: A prospective randomized clinical study was conducted among 40 patients requiring surgical removal of impacted mandibular third molars. All the impacted molars were surgically removed, and the patients were administered bromelain along with amoxicillin. Patients were recalled on day 1, day 3, and day 7 for postoperative pain assessment using visual analogue scale (VAS) and postoperative swelling assessment using facial reference points tragus and pogonion...
December 2016: Journal of International Society of Preventive & Community Dentistry
https://www.readbyqxmd.com/read/28215403/performance-comparison-of-three-trypsin-columns-used-in-liquid-chromatography
#9
Tereza Šlechtová, Martin Gilar, Květa Kalíková, Stephanie M Moore, James W Jorgenson, Eva Tesařová
Trypsin is the most widely used enzyme in proteomic research due to its high specificity. Although the in-solution digestion is predominantly used, it has several drawbacks, such as long digestion times, autolysis, and intolerance to high temperatures or organic solvents. To overcome these shortcomings trypsin was covalently immobilized on solid support and tested for its proteolytic activity. Trypsin was immobilized on bridge-ethyl hybrid silica sorbent with 300Å pores, packed in 2.1×30mm column and compared with Perfinity and Poroszyme trypsin columns...
February 14, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28215327/enzymes-in-fermented-fish
#10
Giyatmi, H E Irianto
Fermented fish products are very popular particularly in Southeast Asian countries. These products have unique characteristics, especially in terms of aroma, flavor, and texture developing during fermentation process. Proteolytic enzymes have a main role in hydrolyzing protein into simpler compounds. Fermentation process of fish relies both on naturally occurring enzymes (in the muscle or the intestinal tract) as well as bacteria. Fermented fish products processed using the whole fish show a different characteristic compared to those prepared from headed and gutted fish...
2017: Advances in Food and Nutrition Research
https://www.readbyqxmd.com/read/28214886/adenovirus-vector-harboring-the-hbcag-and-tripeptidyl-peptidase-ii-genes-induces-potent-cellular-immune-responses-in-vivo
#11
Quanhui Tan, Siyuan Ma, Jianjun Hu, Xiaohua Chen, Yongsheng Yu, Zhenghao Tang, Guoqin Zang
BACKGROUND: Chronic hepatitis B virus (HBV) infection is associated with a weak but specific cellular immune response of the host to HBV. Tripeptidyl peptidaseⅡ (TPPⅡ), an intracellular macromolecule and proteolytic enzyme, plays an important complementary and compensatory role for the proteasome during viral protein degradation and major histocompatibility complex class I antigen presentation by inducing a specific cellular immune response in vivo. Based on a previous study, we aimed to explore the role of MHC class I antigen presentation in vivo and the mechanisms that may be involved...
January 27, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28213441/secreted-metalloproteinase-adamts-3-inactivates-reelin
#12
Himari Ogino, Arisa Hisanaga, Takao Kohno, Yuta Kondo, Kyoko Okumura, Takana Kamei, Tempei Sato, Hiroshi Asahara, Hitomi Tsuiji, Masaki Fukata, Mitsuharu Hattori
The secreted glycoprotein Reelin regulates embryonic brain development and adult brain functions. It has been suggested that reduced Reelin activity contributes to the pathogenesis of several neuropsychiatric and neurodegenerative disorders, such as schizophrenia and Alzheimer's disease; however, non-invasive methods that can upregulate Reelin activity in vivo have yet to be developed. We previously found that the proteolytic cleavage of Reelin within Reelin repeat 3 (N-t site) abolishes Reelin activity in vitro, but it remains controversial as to whether this effect occurs in vivo Here we partially purified the enzyme that mediates the N-t cleavage of Reelin from the culture supernatant of cerebral cortical neurons...
February 17, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28210916/a-multiscale-mathematical-model-of-tumour-invasive-growth
#13
Lu Peng, Dumitru Trucu, Ping Lin, Alastair Thompson, Mark A J Chaplain
Known as one of the hallmarks of cancer (Hanahan and Weinberg in Cell 100:57-70, 2000) cancer cell invasion of human body tissue is a complicated spatio-temporal multiscale process which enables a localised solid tumour to transform into a systemic, metastatic and fatal disease. This process explores and takes advantage of the reciprocal relation that solid tumours establish with the extracellular matrix (ECM) components and other multiple distinct cell types from the surrounding microenvironment. Through the secretion of various proteolytic enzymes such as matrix metalloproteinases or the urokinase plasminogen activator (uPA), the cancer cell population alters the configuration of the surrounding ECM composition and overcomes the physical barriers to ultimately achieve local cancer spread into the surrounding tissue...
February 16, 2017: Bulletin of Mathematical Biology
https://www.readbyqxmd.com/read/28207780/analyzing-pepsin-degradation-assay-conditions-used-for-allergenicity-assessments-to-ensure-that-pepsin-susceptible-and-pepsin-resistant-dietary-proteins-are-distinguishable
#14
Rong Wang, Thomas C Edrington, S Bradley Storrs, Kathleen S Crowley, Jason M Ward, Thomas C Lee, Zi L Liu, Bin Li, Kevin C Glenn
The susceptibility of a dietary protein to proteolytic degradation by digestive enzymes, such as gastric pepsin, provides information on the likelihood of systemic exposure to a structurally intact and biologically active macromolecule, thus informing on the safety of proteins for human and animal consumption. Therefore, the purpose of standardized in vitro degradation studies that are performed during protein safety assessments is to distinguish whether proteins of interest are susceptible or resistant to pepsin degradation via a study design that enables study-to-study comparison...
2017: PloS One
https://www.readbyqxmd.com/read/28207246/aspartate-glycosylation-triggers-isomerization-to-isoaspartate
#15
John Janetzko, Suzanne Walker
O-Linked β-N-acetylglucosamine transferase (OGT) is an essential human enzyme that glycosylates numerous nuclear and cytoplasmic proteins on serine and threonine. It also cleaves Host cell factor 1 (HCF-1) by a mechanism in which the first step involves glycosylation on glutamate. Replacing glutamate with aspartate in an HCF-1 proteolytic repeat was shown to prevent peptide backbone cleavage, but whether aspartate glycosylation occurred was not examined. We report here that OGT glycosylates aspartate much faster than it glycosylates glutamate in an otherwise identical model peptide substrate; moreover, once formed, the glycosyl aspartate reacts further to form a succinimide intermediate that hydrolyzes to produce the corresponding isoaspartyl peptide...
February 21, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28202335/purification-and-characterization-of-a-milk-clotting-aspartic-protease-from-withania-coagulans-fruit
#16
Mahmoud Salehi, Mahmoud Reza Aghamaali, Reza H Sajedi, S Mohsen Asghari, Eisa Jorjani
Withania coagulans fruit has traditionally been used as milk coagulant. The present study reports the purification and characterization of an aspartic protease from W. coagulans fruit. The enzyme was purified via fractional ammonium sulfate precipitation and cation exchange chromatography. SDS-PAGE analysis revealed the presence of a monomeric protein with molecular weight of 31kDa. Proteolytic activity (PA) of the protease was evaluated using casein, and the milk-clotting activity (MCA) was analyzed by skim milk...
February 13, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28199757/genetic-enzymatic-and-metabolite-profiling-of-the-lactobacillus-casei-group-reveals-strain-biodiversity-and-potential-applications-for-flavour-diversification
#17
Ewelina Stefanovic, Kieran N Kilcawley, Mary C Rea, Gerald F Fitzgerald, Olivia McAuliffe
AIMS: The Lactobacillus casei group represents a widely explored group of lactic acid bacteria, characterised by a high level of biodiversity. In this study, the genetic and phenotypic diversity of a collection of more than 300 isolates of the L. casei group and their potential to produce volatile metabolites important for flavour development in dairy products was examined. METHODS AND RESULTS: Following confirmation of species by 16S rRNA PCR, the diversity of the isolates was determined by pulsed field gel electrophoresis...
February 15, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28199445/plant-arginyltransferases-ates
#18
Tatiana Domitrovic, Anna K Fausto, Tatiane da F Silva, Elisson Romanel, Maite F S Vaslin
Regulation of protein stability and/or degradation of misfolded and damaged proteins are essential cellular processes. A part of this regulation is mediated by the so-called N-end rule proteolytic pathway, which, in concert with the ubiquitin proteasome system (UPS), drives protein degradation depending on the N-terminal amino acid sequence. One important enzyme involved in this process is arginyl-t-RNA transferase, known as ATE. This enzyme acts post-translationally by introducing an arginine residue at the N-terminus of specific protein targets to signal degradation via the UPS...
February 13, 2017: Genetics and Molecular Biology
https://www.readbyqxmd.com/read/28196869/structural-insights-unravel-the-zymogenic-mechanism-of-the-virulence-factor-gingipain-k-from-porphyromonas-gingivalis-a-causative-agent-of-gum-disease-from-the-human-oral-microbiome
#19
Anja Pomowski, Isabel Uson, Zuzanna M Nowakowska, Florian Veillard, Maryta N Sztukowska, Tibisay Guevara, Theodoros Goulas, Danuta Mizgalska, Magdalena L Nowak, Barbara A Potempa, James A Huntington, Jan Potempa, F Xavier Gomis-Ruth
Skewing of the human oral microbiome causes dysbiosis and preponderance of bacteria such as Porphyromonas gingivalis, the main etiological agent of periodontitis. P. gingivalis secretes proteolytic gingipains (Kgp and RgpA/B) as zymogens inhibited by a pro-domain that is removed during extracellular activation. Unraveling the molecular mechanism of Kgp zymogenicity is essential to design inhibitors blocking its activity. Here, we found that the isolated 209-residue Kgp pro-domain is a boomerang-shaped all-β protein similar to the RgpB pro-domain...
February 14, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28196299/activity-based-probes-for-the-ubiquitin-conjugation-deconjugation-machinery-new-chemistries-new-tools-and-new-insights
#20
REVIEW
David S Hewings, John A Flygare, Matthew Bogyo, Ingrid E Wertz
The reversible post-translational modification of proteins by ubiquitin and ubiquitin-like proteins regulates almost all cellular processes, by affecting protein degradation, localization, and complex formation. Deubiquitinases (DUBs) are proteases that remove ubiquitin modifications or cleave ubiquitin chains. Most DUBs are cysteine proteases, which makes them well suited for study by activity-based probes. These DUB probes report on deubiquitinase activity by reacting covalently with the active site in an enzyme-catalyzed manner...
February 14, 2017: FEBS Journal
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