Search results for "Protease"

2020

The facile synthesis and detailed investigation of a class of highly potent protease inhibitors based on 1,4-naphthoquinones with a dipeptidic recognition motif (HN-l-Phe-l-Leu-OR) in the 2-position and an electron-withdrawing group (EWG) in the 3-position is presented. One of the compound representatives, namely the acid with EWG = CN and with R = H proved to be a highly potent rhodesain inhibitor with nanomolar affinity. The respective benzyl ester (R = Bn) was found to be hydrolyzed by the target enzyme itself yielding the free acid. Detailed kinetic and mass spectrometry studies revealed a reversible covalent binding mode. Theoretical calculations with different density functionals (DFT…

Two proteases from nuclei of rat testis cells. I. Isolation

1987

Abstract Two proteases, assayed with fluorogenic peptides and tentatively designated Rc and Kc, have been isolated from nuclei of rat testis cells by differential extraction with acetic acid, removal of some proteins at pH 4.5, and polyacrylamide gel electrophoresis followed by electroblotting onto nitrocellulose paper. Protease R hydrolyzes t‐Butyl‐oxycarbonyl‐Val‐Pro‐Arg‐7‐amino‐4‐methyl‐coumarin and other peptides in which arginine is joined to 7‐amino‐4‐methyl‐coumarin by amide linkage. Protease Kc has a preference for peptides terminating in lysine‐7‐amino‐4‐methylcoumarin amide. Neither of these proteases is active against Glu‐Phe‐7‐amino‐4‐methyl‐coumarin amide or Carbobenzoxy‐Arg‐7‐…

Identification, Characterization and Synthesis of Natural Parasitic Cysteine Protease Inhibitors – More Potent Falcitidin Analogs

2021

ABSTRACTProtease inhibitors represent a promising therapeutic option for the treatment of parasitic diseases such as malaria and human African trypanosomiasis. Falcitidin was the first member of a new class of inhibitors of falcipain-2, a cysteine protease of the malaria parasite Plasmodium falciparum. Using a metabolomics dataset of 25 Chitinophaga strains for molecular networking enabled identification of over 30 natural analogs of falcitidin. Based on MS/MS spectra, they vary in their amino acid chain length, sequence, acyl residue, and C-terminal functionalization; therefore, they were grouped into the four falcitidin peptide families A-D. The isolation, characterization and absolute st…

Biotechnological characterisation of exocellular proteases produced by enologicalHanseniasporaisolates

2014

Summary Twenty-six enological Hanseniaspora isolates were identified by ITS PCR-RFLP and D1/D2 domain of 26S rDNA sequencing and assayed for exocellular protease production. Based on qualitative data, six isolates, belonging to H. guilliermondii, H. valbyensis and H. occidentalis species, were selected to continue the study. Analytical procedure was optimised, and protease activities were quantified and characterised on the basis of different biotechnological factors. Protease activity was quite glucose, fructose and ethanol content independent, but divalent cation affects activity; these data support that they were aspartic proteases. The effect of 2-mercaptoethanol suggests the importance…

Two proteases from nuclei of rat testis cells. II. Inhibitors

1987

Abstract Proteases Rc and Kc of the nuclei of rat testis cells exhibit considerable differences in susceptibility to various protease inhibitors. Protease Rc is inhibited by D‐phenylalanyl‐L‐propyl‐L‐arginine chlormethylketone and p‐nitrophenyl‐p1‐guanidino ben‐zoate. Protease Kc is inhibited by Nα‐p‐tosyl‐L‐lysine chlormethylketone, N‐p‐tosyl‐phenylalanine chlormethylketone, N‐carbobenzoxy‐L‐phenylalanine chloromethylketone and p‐nitrophenyl‐p'‐guanidino‐benzoate. Neither protease is inhibited by 10 mM phenylmethanesulphonyl‐fluoride or p‐chloromercuri‐benzoate. p‐nitrophenyl‐p1‐guanidino‐benzoate is a substrate for protease Rc with release of p‐nitrophenol, however the protease activity i…

Receptor for advanced glycation end products is subjected to protein ectodomain shedding by metalloproteinases.

2008

The receptor for advanced glycation end products (RAGE) is a 55-kDa type I membrane glycoprotein of the immunoglobulin superfamily. Ligand-induced up-regulation of RAGE is involved in various pathophysiological processes, including late diabetic complications and Alzheimer disease. Application of recombinant soluble RAGE has been shown to block RAGE-mediated pathophysiological conditions. After expression of full-length RAGE in HEK cells we identified a 48-kDa soluble RAGE form (sRAGE) in the culture medium. This variant of RAGE is smaller than a 51-kDa soluble version derived from alternative splicing. The release of sRAGE can be induced by the phorbol ester PMA and the calcium ionophore c…

Melatonin stimulates the nonamyloidogenic processing ofβAPP through the positive transcriptional regulation of ADAM10 and ADAM17

2014

Melatonin controls many physiological functions including regulation of the circadian rhythm and clearance of free radicals and neuroprotection. Importantly, melatonin levels strongly decrease as we age and patients with Alzheimer's disease (AD) display lower melatonin than age-matched controls. Several studies have reported that melatonin can reduce aggregation and toxicity of amyloid-β peptides that are produced from the β-amyloid precursor protein (βAPP). However, whether melatonin can directly regulate the βAPP-cleaving proteases ('secretases') has not been investigated so far. In this study, we establish that melatonin stimulates the α-secretase cleavage of βAPP in cultured neuronal an…

A new method to value efficiency of enzyme blends for pancreatic tissue digestion.

2010

Islet transplantation, since the 90’s, has been resulting to be one of the best successful example of human cell therapy. Nevertheless, islet isolation procedure is not completely standardized; in fact, more than fifty percent of islets procedures don’t arrive to their transplantation. This is due both to the variability of donor’s pancreas and to an unpredictable enzymatic blend efficiency. Enzymes used in pancreas digestion are extracted from Clostridium histolyticum bacteria and digest several substrates. In particular they have strong collagenolytic activity compared to vertebrate collagenases. However, several impediments persist in human islet isolation success probably due to the var…

Effect of digitonin on membrane-bound and chitosomal chitin synthetase activity in protoplasts from yeast cells ofCandida albicans

1993

The effect of digitonin on chitin synthetase present in membrane (MMF) and cytoplasmic fractions (chitosomes) (CF) from C. albicans yeast protoplasts has been determined. The zymogen is preferentially, but not exclusively, solubilized by digitonin from MMF. Centrifugation of distinct solubilized preparations, containing either zymogen, in vivo active enzyme and/or trypsin activated enzyme, on linear sucrose gradients suggests that both zymogen and trypsin activated enzyme sediment slightly slower than the active enzyme, pointing out differences between the activation processes in vivo and in vitro or, alternatively, that both enzyme activities (active in vivo and zymogenic) correspond to di…

Human Enterovirus Group B Viruses Rely on Vimentin Dynamics for Efficient Processing of Viral Nonstructural Proteins.

2019

A virus needs the host cell in order to replicate and produce new progeny viruses. For this, the virus takes over the host cell and modifies it to become a factory for viral proteins. Irrespective of the specific virus family, these proteins can be divided into structural and nonstructural proteins. Structural proteins are the building blocks for the new progeny virions, whereas the nonstructural proteins orchestrate the takeover of the host cell and its functions. Here, we have shown a mechanism that viruses exploit in order to regulate the host cell. We show that viral protein synthesis induces vimentin cages, which promote production of specific viral proteins that eventually control apo…