• MeSH
• aminopeptidasy analýza   metabolismus   MeSH
• cévy enzymologie   MeSH
• histocytochemie metody   MeSH
• krysa rodu rattus MeSH
• mozek enzymologie   metabolismus   MeSH
• neuroglie enzymologie   MeSH
• neurony enzymologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Schistosoma blood flukes are causative agents of schistosomiasis, a parasitic disease that is a major global health problem with 230 million people infected. Recently, digestive proteolytic enzymes of the blood flukes were evaluated as promising therapeutic targets for the treatment of schistosomiasis. The project focuses on of Schistosoma mansoni through inhibition of two cysteine exopeptidases critical for parasite digestion. First, selective peptidomimetic inhibitors of these exopeptidases will be developed by structure-based rational design and synthesized. Second, the inhibitors will be subjected to a multistep screening including testing in vitro against recombinant enzymes, ex vivo against live parasites in culture, and in vivo in murine model of schistosomiasis. The project will provide new lead drug compounds for anti-schistosomal chemotherapy.

Krevničky rodu Schistosoma jsou původcem schistosomózy, parazitárního infekčního onemocnění, které postihuje 230 miliónů lidí a představuje tak globální zdravotní problém. Trávicí proteolytické enzymy krevničky byly nedávno identifikovány jako nadějné molekulární terapeutické cíle pro léčbu schistosózy. Projekt se zaměřuje na potlačení životaschopnosti krevničky střevní Schistosoma mansoni prostřednictvím inhibice dvou cysteinových exopeptidas, které jsou kritické pro trávení krevniček. Zaprvé, selektivní peptidomimetické inhibitory těchto exopeptidas budou racionálně navrženy s využitím strukturních modelů a syntetizovány. Zadruhé, navržené inhibitory budou funkčně charakterizovány pomocí testovány in vitro s rekombinantními exopeptidasami, ex vivo s živými parazity a in vivo v myším modelu schistosomózy. Projekt přinese nové inhibiční molekuly jako potenciální antischistosomální léčiva.

• Klíčová slova
• krevnička, Schistosoma, cysteinová exopeptidasa, terapeutický cíl, návrh a syntéza inhibitorů, schistosomóza, léčiva proti krevničce, blood fluke, Schistosoma, cysteine exopeptidase, drug target, design and synthesis of inhibitors, schistosomiasis, antischistosomal chemotherapeutics,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

• MeSH
• exopeptidasy izolace a purifikace   MeSH
• klinické enzymatické testy MeSH
• LD50 MeSH
• Serratia marcescens enzymologie   klasifikace   patogenita   MeSH
• virulence MeSH

• MeSH
• Candida izolace a purifikace   patogenita   MeSH
• exopeptidasy MeSH
• faktory virulence MeSH
• finanční podpora výzkumu jako téma MeSH

• MeSH
• Alzheimerova nemoc etiologie   mikrobiologie   terapie   MeSH
• bakteriální proteiny metabolismus   MeSH
• exopeptidasy analýza   farmakologie   MeSH
• nádory etiologie   mikrobiologie   terapie   MeSH
• Streptococcus enzymologie   MeSH

• MeSH
• akutní poškození ledvin etiologie   MeSH
• diseminovaná intravaskulární koagulace komplikace   MeSH
• exopeptidasy genetika   klasifikace   nedostatek   MeSH
• lidé MeSH
• syndrom systémové zánětlivé reakce komplikace   patofyziologie   MeSH
• trombotická trombocytopenická purpura diagnóza   patofyziologie   MeSH
• von Willebrandův faktor metabolismus   škodlivé účinky   MeSH
• Check Tag
• lidé MeSH

The accumulation of amyloid-β (Aβ) peptide is thought to be a major causative mechanism of Alzheimer's disease. Aβ accumulation could be caused by dysregulated processing of amyloid precursor protein, yielding excessive amounts of Aβ, and/or by inefficient proteolytic degradation of the peptide itself. Several proteases have been described as Aβ degradation enzymes, most notably metalloendopeptidases, aspartic endopeptidases, and some exopeptidases. Recently a report suggested that another metallopeptidase, glutamate carboxypeptidase II (GCPII), can also cleave Aβ. GCPII is a zinc exopeptidase that cleaves glutamate from N-acetyl-L-aspartyl-L-glutamate in the central nervous system and from pteroylpoly-γ-glutamate in the jejunum. GCPII has been proposed as a promising therapeutic target for disorders caused by glutamate neurotoxicity. However, an Aβ-degrading activity of GCPII would compromise potential pharmaceutical use of GCPII inhibitors, because the enzyme inhibition might lead to increased Aβ levels and consequently to Alzheimer's disease. Therefore, we analyzed the reported Aβ-degrading activity of GCPII using highly purified recombinant enzyme and synthetic Aβ. We did not detect any Aβ degradation activity of GCPII or its homologue even under prolonged incubation at a high enzyme to substrate ratio. These results are in good agreement with the current detailed structural understanding of the substrate specificity and enzyme-ligand interactions of GCPII.

• MeSH
• amyloidní beta-protein chemie   metabolismus   MeSH
• antigeny povrchové genetika   metabolismus   MeSH
• biokatalýza účinky léků   MeSH
• dipeptidy metabolismus   MeSH
• glutamátkarboxypeptidasa II antagonisté a inhibitory   genetika   metabolismus   MeSH
• hmotnostní spektrometrie MeSH
• hydrolýza MeSH
• katalytická doména MeSH
• lidé MeSH
• molekulární struktura MeSH
• neprilysin genetika   metabolismus   MeSH
• organofosforové sloučeniny farmakologie   MeSH
• peptidové fragmenty chemie   metabolismus   MeSH
• proteolýza MeSH
• rekombinantní proteiny metabolismus   MeSH
• substrátová specifita MeSH
• tritium MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Glutamate carboxypeptidase II (GCPII) is a transmembrane enzyme that cleaves N-acetyl-L-aspartyl-L-glutamate (NAAG) in the brain. GCPII is highly expressed in the prostate and prostate cancer and might be associated with prostate cancer progression. Another exopeptidase, plasma glutamate carboxypeptidase (PGCP), was reported to be similar to GCPII and to share its NAAG-hydrolyzing activity. METHODS: We performed a radioenzymatic assay with [(3) H]NAAG as a substrate to detect and quantify the enzymatic activity of GCPII in plasma. Using a specific antibody raised against native GCPII (2G7), we immunoprecipitated GCPII from human plasma. We also cloned two PGCP constructs, expressed them in insect cells, and tested them for their NAAG-hydrolyzing activity. RESULTS: We detected GCPII protein in human plasma and found that its concentration ranges between 1.3 and 17.2 ng/ml in volunteers not diagnosed with prostate cancer. Recombinant PGCP was enzymatically active but exhibited no NAAG-hydrolyzing activity. CONCLUSION: GCPII is present in human blood, and its concentration within a healthy population varies. Recombinant PGCP does not hydrolyze NAAG, suggesting that GCPII alone is responsible for the NAAG-hydrolyzing activity observed in human blood. The potential correlation between GCPII serum levels and the disease status of prostate cancer patients will be further investigated.

• MeSH
• dospělí MeSH
• glutamátkarboxypeptidasa II krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery krev   MeSH
• nádory prostaty krev   diagnóza   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Host blood proteins, represented mainly by hemoglobin and serum albumin, serve as the ultimate source of amino acids needed for de novo protein synthesis during tick development and reproduction. While uptake and processing of hemoglobin by tick gut cells have been studied in detail, molecular mechanisms of host serum albumin degradation remain unknown. In this work, we have used artificial membrane feeding of Ixodes ricinus females on a hemoglobin-free diet in order to characterize the proteolytic machinery involved in albuminolysis. Morphological comparisons of ticks fed on whole blood (BF) and serum (SF) at microscopic and ultrastructural levels showed that albumin and hemoglobin have different trafficking routes in tick gut cells. Analysis in vitro with selective inhibitors demonstrated that albumin is degraded at an acidic pH by a network of cysteine and aspartic peptidases with predominant involvement of cysteine cathepsins having endo- and exopeptidase activities. The cleavage map of albumin and the roles of individual peptidases in albumin degradation were determined. These results indicate that the albuminolytic pathway is controlled by the same proteolytic system that is responsible for hemoglobinolysis. This was further supported by the overall similarity of gut peptidase profiles in SF and BF ticks at the transcriptional and enzymatic activity levels. In conclusion, our work provides evidence that although hemoglobin and albumin are transported differentially during heterophagy they are digested by a common multienzyme proteolytic network. This central digestive system, critical for successful blood feeding in tick females, thus represents a valuable target for novel anti-tick interventions.

• MeSH
• aspartátové proteasy metabolismus   MeSH
• cysteinové proteasy metabolismus   MeSH
• hemoglobiny metabolismus   MeSH
• klíště enzymologie   MeSH
• koncentrace vodíkových iontů MeSH
• proteolýza * MeSH
• sérový albumin metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

There is a new public health crisis threatening the world with the emergence and spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The disease was later named novel coronavirus disease or COVID-19. It was then declared a pandemic by the World Health Organization on March 11, 2020. The virus originated in bats and was transmitted to humans through unknown intermediary animals in Wuhan, Hubei province, China, in December 2019. As of February 5, 2021, 103 million laboratory-confirmed cases and nearly 2.3 million deaths were reported globally. The number of death tolls continues to rise, and a large number of countries have been forced to maintain social distance in public place and enforce lockdown. As per literature, coronavirus is transmitted human to human or human to animal via airborne droplets. Coronavirus enters the human cell through the membrane ACE-2 exopeptidase receptor. WHO, ECDC, and ICMR advised avoiding public places and close contact with infected persons and pet animals. To date, there is no evidence of any effective treatment for COVID-19. The main therapies being used to treat the disease are antiviral drugs, chloroquine/hydroxychloroquine, and respiratory therapy. Although several therapies have been proposed, quarantine is the only intervention that appears to be effective in decreasing the contagion rate. We conducted a literature review of publicly available information to summarize knowledge about the pathogen and the current epidemic. In the present literature review, the causative agent of the pandemic, epidemiology, pathogenesis, and diagnostic techniques are discussed. Further, currently used treatment, preventive strategies along with vaccine trials and computational tools are all described in detail.

• MeSH
• COVID-19 * MeSH
• hydroxychlorochin MeSH
• kontrola infekčních nemocí MeSH
• lidé MeSH
• pandemie MeSH
• SARS-CoV-2 * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH