RESUMO
The antibacterial activity of BK-218 was similar to that of cefamandole when it was tested against several laboratory strains. The inhibiting effect of BK-218 was greater than that of cephalexin and cefoxitin on penicillin-binding proteins of Escherichia coli HB101. This result was in close correlation with the relative inhibition of radiolabeled glucosamine incorporation (greatest with BK-218) and with the lytic effect (most intensive with BK-218). BK-218 proved to be a good inhibitor for all five of the beta-lactamases that were investigated, although two enzymes (Enterobacter cloacae P99 and Pseudomonas aeruginosa Cilote) hydrolyzed it to some extent.
Assuntos
Bactérias/efeitos dos fármacos , Proteínas de Bactérias , Cefalosporinas/farmacologia , Hexosiltransferases , Peptidil Transferases , Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Parede Celular/metabolismo , Cefalosporinas/metabolismo , Cefalosporinas/farmacocinética , Citoplasma/metabolismo , Enterococcus faecalis/metabolismo , Escherichia coli/metabolismo , Glucosamina/metabolismo , Testes de Sensibilidade Microbiana , Muramilpentapeptídeo Carboxipeptidase/metabolismo , Proteínas de Ligação às Penicilinas , beta-Lactamases/metabolismoAssuntos
Catepsinas/sangue , Glicosaminoglicanos/metabolismo , Granulócitos/enzimologia , Elastase Pancreática/sangue , Catepsina G , Sulfatos de Condroitina/metabolismo , Cromatografia de Afinidade , Dermatan Sulfato/metabolismo , Heparina/metabolismo , Heparitina Sulfato/metabolismo , Humanos , Ácido Hialurônico/metabolismo , Serina EndopeptidasesRESUMO
The extracts of granules isolated from bovine granulocytes show elastase- and chymotrypsin-like activities, as detected with specific synthetic substrates. Extraction of these enzymes depends upon salt concentration. In the course of the present studies a 21-fold purification of the elastase-like enzyme was achieved on a (Ala)3-CH-Sepharose 4B gel. The molecular weight of the enzyme is 33 000, as determined by gel electrophoresis in the presence of sodium dodecyl sulfate. The elastase-like activity is inhibited by phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, basic pancreatic inhibitor and by heparin at different rates. Elastatinal inhibits the enzyme competitively (Ki = 80 microM). The cytosol of bovine granulocytes contains a protein which strongly inhibits the elastase-like enzyme of the bovine granulocyte (Ki = 0.4 nM) as well as porcine pancreatic elastase (Ki = 11 nM).
Assuntos
Granulócitos/enzimologia , Elastase Pancreática/isolamento & purificação , Animais , Bovinos , Cromatografia de Afinidade , Citosol/metabolismo , Heparina/farmacologia , Oligopeptídeos/farmacologia , Elastase Pancreática/antagonistas & inibidores , Fluoreto de Fenilmetilsulfonil/farmacologia , Inibidores de Proteases/metabolismo , Suínos , Inibidor da Tripsina de Soja de Bowman-Birk/farmacologia , Inibidor da Tripsina de Soja de Kunitz/farmacologiaRESUMO
The effect of human granulocyte proteases and related pancreatic enzymes on the procoagulant (VIII:C) and the von Wille-brand (VIIIR:WF) activities of factor VIII was investigated. VIII:C appeared to be several hundred times more sensitive towards granulocyute enzymes than VIIIR:WF. The elastase-like protease (ELP) decreased both activities of factor VIII more effectively than the chymotrypsin-like enzyme (CLP). ELP increased co-operatively the inactivating effect of CLP. In the presence of a CLP-ELP mixture in a ratio of 3:1, the rate of inactivation of VIII:C was seven times as high as that with CLP alone. Factor VIII was more resistant towards the damaging effect of pancreatic enzymes. The rate of inactivation of VIII:C was about a thousand times and that of VIIIR:WF about a hundred times lowere than those measured with the related granulocyte enzymes of the same proteolytic activity. The sensitivity of VIII:C towards proteolysis was not as pronounced with pancreatic enzymes as with granulocyte proteases. Our data suggest that, even though CLP and ELP are called "-like# enzymes, the specificity of granulocyte proteases is not identical with that of the pancreatic enzymes. The extreme sensitivity of VIII:C for granulocyte proteases may bring about blood coagulation disorders in certain pathological conditions.
