ABSTRACT
Pemphigus vulgaris is a severe, socially significant autoimmune disease associated with autoantibodies to the desmoglein 3 antigen. The disease affects all age groups, beginning at 18 years of age; the mortality rate of pemphigus can reach as high as 50%, depending on a patient's age and a number of other factors. There is no highly selective or personalized therapy for pemphigus vulgaris at the moment. One of the well-known therapeutic approaches to the disease is to use rituximab, an anti-CD20 antibody that can help achieve B cell depletion in peripheral blood. To solve the problem of nonspecific elimination of B cells in patients with pemphigus vulgaris, it is reasonable to use specific immunoligands, their choice being based on an assessment of the level of autoantibodies specific to each of the fragments of desmoglein. In this work, the proportion of autoreactive B cells in patients diagnosed with pemphigus vulgaris is found to be 0.09-0.16%; a positive correlation was revealed between the antibody level and the number of autoreactive B cells to various fragments of desmoglein.
Subject(s)
Drug Resistance, Multiple/genetics , HIV Protease Inhibitors/pharmacology , HIV Protease/genetics , Indinavir/pharmacology , Molecular Dynamics Simulation , Mutation/genetics , Sulfonamides/pharmacology , Amino Acid Substitution/genetics , Darunavir , Drug Design , Fluorescent Dyes/pharmacology , HIV Protease/chemistry , HIV Protease/metabolism , Humans , KineticsABSTRACT
A thermophilic anaerobic bacterial strain 1004-09 belonging to the genus Thermoanaerobacter and capable of growth on protein substrates such as albumin, gelatin, casein, and alpha and beta-keratins was isolated from the Urinskii hot spring (Barguzin river valley, Republic of Buryatia, Russia). A 150-kDa serine proteinase was revealed in the strain supernatant; it exhibited optimal activity at 60 degrees C and pH 9.3 and was capable of keratin hydrolysis. A number of characteristics for the strain 1004-09 keratinase were established including activation by SDS and NaCl and residual activity (15% to the activity of the intact protein) in the presence of 10% ethanol and acetone.
Subject(s)
Hot Springs/microbiology , Peptide Hydrolases/metabolism , Thermoanaerobacter/enzymology , Water Microbiology , Enzyme Activators/pharmacology , Hydrogen-Ion Concentration , Hydrolysis , Keratins/metabolism , Molecular Weight , Peptide Hydrolases/chemistry , Peptide Hydrolases/drug effects , Siberia , Sodium Chloride/pharmacology , Sodium Dodecyl Sulfate/pharmacology , Temperature , Thermoanaerobacter/classificationABSTRACT
A number of new inhibitors of plasmepsin II (PlmII) from Plasmodium falciparum, one of the key factors of malarial parasite survival, were synthesized. The inhibitors are analogues of pepstatin with various variants of Ala residue substitutions. Effects of the inhibitors on human PlmII and cathepsin D were studied. Inhibition of PlmII by the substrate was found, which required the use of the modified Henderson method for the determination of inhibition constants. Two synthesized inhibitors were shown to exhibit a pronounced selectivity to PlmII (K(i) = 5.5 and 5 nM) in comparison with cathepsin D (K(i) = 230 and 3000 nM, respectively).
Subject(s)
Aspartic Acid Endopeptidases/antagonists & inhibitors , Plasmodium falciparum/enzymology , Protease Inhibitors/chemistry , Protozoan Proteins/antagonists & inhibitors , Animals , Aspartic Acid Endopeptidases/chemistry , Cathepsin D/antagonists & inhibitors , Cathepsin D/chemistry , Humans , Pepstatins/chemistry , Protozoan Proteins/chemistryABSTRACT
Target-aimed synthesis of a new class of water soluble amino acid and dipeptide derivatives of fullurene (C60 - X) for inhibition of specific virus enzymes, i.e. protease and reverse transcriptase of HIV (P HIV and RT HIV) in cell culture lytic and chronic infections was performed. Out of 13 tested substances, 8 showed inhibitory activity and 5 were effective in pharmacological doses (ID50 varied within 0.46 to 1.0 mcm/ml with respect to the lytic infection and 5.0 to 12.5 mcm/ml with respect to the chronic infection). The activity of (1), (2), (6), (7) and (8) was comparable to that of azidothymidine, a nucleozide inhibitor of RT HIV in the cell culture lytic infection. The substances also showed marked virucidal action. The cytotoxicity (survival, antiproliferative effect) varied from low to very low with respect to the rapidly dividing cells MT4 and HTHIV27 (CD50 > 200-800) and was somewhat higher with respect to PBL (CD50 > 100). The selectivity index (SI = CD50/ID50) was equal to 165-2000 for various samples. The prototype derivatives (1) and (2) had a selective (competitive) inhibitory action on the recombinant protease of HIV with IC50 = 1.25-2.76 mcM, while derivatives (1), (la) and (2) had a noncompetitive inhibitory action on the recombinant reverse transcriptase of HIV (Ki = 7.9-12.1 mM). The pharmacokinetic study of the prototype derivative (1) on laboratory animals revealed no acute or chronic toxicity up to the terminal high concentrations. As for (1), its high interspecies (mice--rabbits) relative bioavailability equal to 110% was shown.
Subject(s)
Fullerenes/pharmacology , HIV Protease Inhibitors/pharmacology , HIV Reverse Transcriptase/antagonists & inhibitors , HIV-1/physiology , Reverse Transcriptase Inhibitors/pharmacology , Virus Replication/drug effects , Amino Acids/chemistry , Amino Acids/pharmacology , Animals , Cell Line , Dipeptides/chemistry , Dipeptides/pharmacology , Dose-Response Relationship, Drug , Fullerenes/chemistry , HIV-1/drug effects , Humans , Mice , Virus Replication/physiologyABSTRACT
Neurochordins constitute a family of several immunologically interrelated nervous tissue glycoproteins occurring in the soluble protein fraction of the brain of all vertebrate species including human. The only exception is neurochordin D which, as it has been shown in the present study, is present in human brain but not in the brains of rats and chickens. Human neurochordin D is first detected in the brain at the age of 18 months and further persists through life. A highly purified neurochordin D preparation has been obtained by immunoaffinity chromatography combined with biochemical protein fractionation. Quantitative carbohydrate analysis revealed the presence of Gal, Man, GlcNAc, Fuc and GalNAc in the neurochordin D preparation. The sensitivity of neurochordin D to neuraminidase suggests the presence of sialic acids.
Subject(s)
Brain/metabolism , Nerve Tissue Proteins/metabolism , Animals , Carbohydrates/analysis , Chickens , Chromatography, Affinity , Gene Expression Regulation, Developmental , Humans , Nerve Tissue Proteins/chemistry , Nerve Tissue Proteins/genetics , RatsABSTRACT
Glycopeptides were obtained by the pepsin (glycononapeptide) and trypsin (glycohexapeptide) hydrolysis of ovalbumin. A method (HPLC) of the glycopeptidamidase activity determination with a fluorescent-labelled (Dns) glycopeptides is described, and kinetic constants of hydrolysis of specific substrates by glycopeptidamidase A are determined. Dns(O-Dns)Tyr-Asn(CHO)-Leu-Thr-Ser-Val is shown to be a specific substrate of the glycopeptidamidase A.
Subject(s)
Amidohydrolases/metabolism , Amino Acid Sequence , Chromatography, High Pressure Liquid , Dansyl Compounds/metabolism , Hydrolysis , Kinetics , Molecular Sequence Data , Oligopeptides/metabolism , Pepsin A/metabolism , Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase , Substrate Specificity , Trypsin/metabolismABSTRACT
Major types of oligosaccharides linked to protein and glycosidases used for carbohydrate cleavage and analysis are reviewed. Approaches to partial or complete separation of the protein and carbohydrate moieties of glycoproteins are suggested. Some date on the chemical deglycosylation are also presented.
