• Klíčová slova
• ANTIBODIES *,
• MeSH
• bioreaktory * MeSH
• fermentace * MeSH
• lidé MeSH
• papain * MeSH
• protilátky * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• papain * MeSH
• protilátky * MeSH

Adsorption properties of protein Papain at the solid|liquid (0.1 M KCl) interfaces of different hydrophobicity [highly oriented pyrolytic graphite (HOPG), bare gold, CH3, OH, and COOH-terminated self-assembled monolayers on gold] were studied by a combined quartz crystal microbalance and atomic force microscopy techniques. It was found that Papain forms an incomplete monolayer at hydrophobic interfaces (HOPG and CH3-terminated substrate), whereas on more hydrophilic ones, a complete monolayer formation was always observed with either the onset of the formation of a second layer (bare gold substrate) or adsorption in a multilayer fashion, possibly a bilayer formation (OH-terminated substrate). The surface concentration and compact monolayer film thickness was much lower on the COOH-terminated substrate compared to other surfaces studied. This result was explained by partial dissociation of the interfacial COOH groups leading to additional electrostatic interactions between the positively charged protein domains and negatively charged carboxylate anions, as well as to local pH changes promoting protein denaturation.

• MeSH
• adsorpce * MeSH
• grafit chemie   MeSH
• hydrofobní a hydrofilní interakce * MeSH
• mikrorovnovážné techniky křemenného krystalu MeSH
• mikroskopie atomárních sil MeSH
• papain metabolismus   MeSH
• povrchové vlastnosti * MeSH
• zlato chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• grafit MeSH
• papain MeSH
• zlato MeSH

Nanobubbles formed on monocrystalline gold/water interface by means of the ethanol-to-water solvent exchange were exposed to the solutions of either bovine serum albumin or papain proteins. Both proteins do not change the position of nanobubbles in water, as observed by in situ tapping mode atomic force microscopy imaging before and after the introduction of the protein. The aqueous environment was subsequently replaced by ethanol. While all nanobubbles were found to dissolve in ethanol in the presence of bovine serum albumin, most of them survived when papain was employed. The protective ability of papain was ascribed to its resistance towards the protein denaturation in aqueous solutions of ethanol. The authors employed in situ atomic force nanolithography to investigate the nanomorphology of the papain/nanobubble assemblies in ethanol.

• MeSH
• ethanol chemie   MeSH
• nanostruktury chemie   MeSH
• papain metabolismus   MeSH
• povrchové vlastnosti MeSH
• sérový albumin hovězí metabolismus   MeSH
• skot MeSH
• voda chemie   MeSH
• zlato metabolismus   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ethanol MeSH
• papain MeSH
• sérový albumin hovězí MeSH
• voda MeSH
• zlato MeSH

• MeSH
• ovoce enzymologie   MeSH
• papain chemie   izolace a purifikace   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Uzbekistán MeSH
• Názvy látek
• papain MeSH

• Klíčová slova
• AORTA *, EXPERIMENTAL LAB STUDY *, GLYCOPROTEINS *, HISTOCYTOCHEMISTRY *, MICE *, MUCOPOLYSACCHARIDES *, PAPAIN *, PHARMACOLOGY *, POLYSACCHARIDES *, RABBITS *,
• MeSH
• aorta * MeSH
• farmakologie * MeSH
• glykoproteiny * MeSH
• glykosaminoglykany * MeSH
• histocytochemie * MeSH
• králíci * MeSH
• myši * MeSH
• papain * MeSH
• polysacharidy * MeSH
• výzkum * MeSH
• Check Tag
• králíci * MeSH
• myši * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glykoproteiny * MeSH
• glykosaminoglykany * MeSH
• papain * MeSH
• polysacharidy * MeSH

The rate of degradation of poly[N5-(2-hydroxyethyl)-L-glutamine] (PHEG), poly(L-glutamic acid) (PGA) and poly[HEG-co-GA] random copolymers by papain was measured in the pH range 4.0-7.5, employing the gel permeation chromatography method. The effect of the degree of ionization on the polymer conformation was measured by circular dichroism (c.d.). PHEG, which is uncharged, had a random coil conformation and an almost constant degradation rate within the whole pH interval. The ionization of PGA increased with increasing pH and was accompanied by conformational transition from helix to random coil. The hydrolysis of PGA by papain depended on pH with the optimum at about pH 5, indicating that both the high content of helix (at pH less than 5) and increasing charge density (at pH greater than 5), decreased the degradation rate. Contrary to PGA, pH profiles of the degradation rate of poly[HEG-co-GA] copolymers are monotonous and do not decrease at pH less than 5. In the copolymers the HEG residues act as a helix breaker and limit the formation of helical conformation. The role of structural features of a macromolecular substrate, i.e. the charge, helical conformation and the nature of amino acid residues, in the interaction between enzyme and polymer is discussed.

• MeSH
• cirkulární dichroismus MeSH
• gelová chromatografie MeSH
• glutamáty chemie   metabolismus   MeSH
• koncentrace vodíkových iontů MeSH
• konformace proteinů MeSH
• kyselina glutamová MeSH
• papain metabolismus   MeSH
• peptidy chemie   metabolismus   MeSH
• polymery MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glutamáty MeSH
• kyselina glutamová MeSH
• papain MeSH
• peptidy MeSH
• poly-N(5)-(2-hydroxyethyl)glutamine MeSH Prohlížeč
• polymery MeSH

The worldwide COVID-19 pandemic caused by the coronavirus SARS-CoV-2 urgently demands novel direct antiviral treatments. The main protease (Mpro) and papain-like protease (PLpro) are attractive drug targets among coronaviruses due to their essential role in processing the polyproteins translated from the viral RNA. In this study, we virtually screened 688 naphthoquinoidal compounds and derivatives against Mpro of SARS-CoV-2. Twenty-four derivatives were selected and evaluated in biochemical assays against Mpro using a novel fluorogenic substrate. In parallel, these compounds were also assayed with SARS-CoV-2 PLpro. Four compounds inhibited Mpro with half-maximal inhibitory concentration (IC50) values between 0.41 μM and 9.0 μM. In addition, three compounds inhibited PLpro with IC50 ranging from 1.9 μM to 3.3 μM. To verify the specificity of Mpro and PLpro inhibitors, our experiments included an assessment of common causes of false positives such as aggregation, high compound fluorescence, and inhibition by enzyme oxidation. Altogether, we confirmed novel classes of specific Mpro and PLpro inhibitors. Molecular dynamics simulations suggest stable binding modes for Mpro inhibitors with frequent interactions with residues in the S1 and S2 pockets of the active site. For two PLpro inhibitors, interactions occur in the S3 and S4 pockets. In summary, our structure-based computational and biochemical approach identified novel naphthoquinonal scaffolds that can be further explored as SARS-CoV-2 antivirals.

• MeSH
• antivirové látky * farmakologie   chemie   MeSH
• COVID-19 MeSH
• inhibitory proteas * farmakologie   chemie   MeSH
• koronavirové proteasy 3C * antagonisté a inhibitory   MeSH
• koronavirové proteasy podobné papainu * antagonisté a inhibitory   MeSH
• lidé MeSH
• naftochinony * chemie   farmakologie   MeSH
• papain MeSH
• SARS-CoV-2 * účinky léků   enzymologie   MeSH
• simulace molekulového dockingu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• 3C-like proteinase, SARS-CoV-2 MeSH Prohlížeč
• antivirové látky * MeSH
• inhibitory proteas * MeSH
• koronavirové proteasy 3C * MeSH
• koronavirové proteasy podobné papainu * MeSH
• naftochinony * MeSH
• papain-like protease, SARS-CoV-2 MeSH Prohlížeč
• papain MeSH

• MeSH
• detergenty * MeSH
• histokompatibilní antigeny * MeSH
• HLA antigeny * MeSH
• isoantigeny analýza   MeSH
• molekulová hmotnost MeSH
• myši MeSH
• papain * MeSH
• rozpustnost MeSH
• tvorba protilátek MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• detergenty * MeSH
• histokompatibilní antigeny * MeSH
• HLA antigeny * MeSH
• isoantigeny MeSH
• papain * MeSH

PURPOSE: The aim of the present study was to investigate the effect of a mixture of proteolytic enzymes (comprising trypsin, chymotrypsin and papain) on the metastatic model of syngeneic melanoma B16. METHODS: 140 C57B16 mice were divided into two control and two "treated" groups. Control groups received saline rectally, twice a day starting 24 h after intracutaneous transplantation (C1) or from the time point of the primary B16 melanoma extirpation (C2), respectively. "Treated" groups were rectally administered a mixture of 0.2 mg trypsin, 0.5 mg papain, and 0.2 mg chymotrypsin twice daily starting 24 h after transplantation (E1) or after extirpation of the tumor (E2), respectively. Survival of mice and B16 melanoma generalization were observed for a period of 100 days. Immunological evaluation of B16 melanoma cells in the ascites was accomplished. CD44, CD54 and CD106 cells were measured by flow cytometry. RESULTS: Administration of proteolytic enzymes to mice inhibited the growth of primary tumors, and tumor recurrences were less numerous. Importantly, metastasis was considerably curtailed both in the vicinity of the primary tumor and at distant locales. These findings correlated with a decreased expression of CD44 and CD54 molecules in tumors exposed to proteolytic enzymes in vivo. CONCLUSIONS: Our data suggest that serine and cysteine proteinases suppress B16 melanoma, and restrict its metastatic dissemination in C57B16 mice.

• MeSH
• antigeny CD44 imunologie   MeSH
• antigeny povrchové imunologie   MeSH
• aplikace rektální MeSH
• buněčné dělení účinky léků   MeSH
• chymotrypsin aplikace a dávkování   farmakologie   MeSH
• endopeptidasy farmakologie   MeSH
• fixní kombinace léků MeSH
• imunoglobuliny imunologie   MeSH
• melanom experimentální farmakoterapie   imunologie   patologie   sekundární   MeSH
• mezibuněčná adhezivní molekula-1 imunologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory plic imunologie   prevence a kontrola   sekundární   MeSH
• papain aplikace a dávkování   farmakologie   MeSH
• protinádorové látky farmakologie   MeSH
• trypsin aplikace a dávkování   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen 106 MeSH Prohlížeč
• antigeny CD44 MeSH
• antigeny povrchové MeSH
• chymotrypsin, papain, trypsin drug combination MeSH Prohlížeč
• chymotrypsin MeSH
• endopeptidasy MeSH
• fixní kombinace léků MeSH
• imunoglobuliny MeSH
• mezibuněčná adhezivní molekula-1 MeSH
• papain MeSH
• protinádorové látky MeSH
• trypsin MeSH

Hyaluronic acid (HA) is known to serve as a dynamic mediator intervening in many physiological functions. Its specific effect has been repeatedly confirmed to be strongly influenced by the molecular size of hyaluronan fragments. However common technological approaches of HA fragments production have their limitations. In many cases, the final products do not meet the strict pharmaceutical requirements, specifically due to size polydispersity and reaction contaminants. We present novel methodology based on combination of unique incidental ability of the plant-derived protease papain to split the glycosidic bonds and an indispensable advantages of biocompatible macroporous material with incorporated ferrous ions serving as carrier for covalent papain fixation. This atypical and yet unpublished highly efficient multiparametric approach allows enhanced HA fragmentation for easily and safely producing molar-mass-defined HA fragments with narrow size distribution. Native polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography/multi-angle light scattering (SEC-MALS) confirmed the effectiveness of our multiparametric approach.

• Klíčová slova
• Hyaluronan fragmentation, Magnetic particles, Papain, Pharmaceutics, SEC-MALS,
• MeSH
• celulosa chemie   MeSH
• enzymy imobilizované chemie   MeSH
• gelová chromatografie MeSH
• hyaluronoglukosaminidasa chemie   MeSH
• kyselina hyaluronová chemie   MeSH
• magnetické jevy MeSH
• molekulová hmotnost MeSH
• nativní elektroforéza na polyakrylamidovém gelu MeSH
• papain chemie   MeSH
• radiační rozptyl MeSH
• viskozita MeSH
• železo chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• celulosa MeSH
• enzymy imobilizované MeSH
• hyaluronoglukosaminidasa MeSH
• kyselina hyaluronová MeSH
• papain MeSH
• železo MeSH