Subject(s)
Antigens/isolation & purification , Collagen/immunology , Animals , Cattle , Epitopes , Hemagglutination Inhibition Tests/methods , Humans , RabbitsSubject(s)
Glycosaminoglycans/urine , Adolescent , Adult , Age Factors , Child, Preschool , Female , Glycosaminoglycans/analysis , Humans , Male , Methods , Mucopolysaccharidoses/urineABSTRACT
1. A neutral proteinase (EC 3.4.-.-) with elastolytic activity was isolated from canine bloodstream leucocytes, and purified to apparent homogeneity by a two-step procedure consisting of DEAE-Sephadex chromatography and molecular sieving on Sephadex G-75. 2. The molecular weight of the enzyme was 23 500, and the absorbance (A1%1cm) at 282 nm was 6.1. Amino acid analysis showed high content of glycine, aspartic acid, and valine, and low proportion of methionine, lysine and histidine as well as the absence of tyrosine in the enzyme molecule. 3. The proteinase was active against several protein substrates as well as towards N-t-butyloxycarbonyl-L-alanine p-nitrophenyl ester, N-acetyl-L-alanyl-tyrosine ethyl ester. 4. The enzyme was inactivated by diisopropylfluorophosphate, N-acetyl-L-alanyl-L-alanyl-L-alanine chloromethyl ketone, and N-p-tosyl-L-phenylalanine chloromethyl ketone. Inhibition by some natural proteinase inhibitors was also noted.
Subject(s)
Leukocytes/enzymology , Pancreatic Elastase/blood , Peptide Hydrolases/blood , Amino Acids/analysis , Animals , Dogs , Hydrogen-Ion Concentration , Kinetics , Pancreatic Elastase/isolation & purification , Pancreatic Elastase/metabolism , Peptide Hydrolases/isolation & purification , Peptide Hydrolases/metabolism , Structure-Activity RelationshipABSTRACT
Production of a factor activating leucocyte procollagenase by tissue culture of rheumatoid synovium was shown. The activator was isolated, partly purified, and shown to be thermolabile, nondialysable, and had no activity toward casein, haemoglobin, histones, and PZ-peptide. The activity of the activator was partly decreased by trypsin.
Subject(s)
Enzyme Precursors/metabolism , Microbial Collagenase/metabolism , Arthritis, Rheumatoid/enzymology , Culture Techniques , Humans , Synovial Fluid/enzymology , ViscosityABSTRACT
1. An activator catalysing specifically conversion of latent forms of human leucocyte collagenase and gelatin-specific protease into the active forms, has been isolated from rheumatoid synovial fluid and purified 55-fold with a yield of 16%. 2. Molecular weight of the activator is about 35 000. 3. The activator is thermolabile, and is irreversibly inactivated at pH below 5.5 or in the presence of low concentrations of trypsin or papain; it is resistant to the action of lysozyme, hyaluronidase, diisopropylfluorophosphate, soybean trypsin inhibitor, p-chloromercuribenzoate, iodoacetamide and dithiothreitol. 4. The activator did not show any activity towards collagen, gelatin, casein, haemoglobin, histones, elastin or p-phenylazobenzyloxycarbonyl-peptide.
