Isolation and characterization of five fibrin(ogen)olytic enzymes from the venom of Philodryas olfersii (green snake).
Five distinct fibrin(ogen)olytic proteinases PofibC1, C2, C3, H and S were isolated by gel filtration and ion-exchange chromatographies. PofibC1, C2, C3 and H are metalloproteinases inhibited by ethylenediamine tetracetic acid (EDTA) or 1,10-phenanthroline. Only PofibH had hemorrhagic activity. PofibS is a serine proteinase, inhibited by phenylmethylsulfonyl fluoride (PMSF) or Torresea cearensis trypsin inhibitor (TCTI). All five enzymes were inhibited by dithiothreitol (DTT) or dithioerythritol (DTE). PofibC1 and C2 presented the same mol. wt of 47,000 and are acidic proteins of pI 6.2 PofibC3 is a basic proteinase of pI 8.5 and mol. wt 45,000. The hemorrhagic proteinase PofibH had a mol. wt of 58,000 and pI of 4.6 and PofibS had a mol. wt of 36,000 and pI of 4.5. The five proteinases degraded fibrin and fibrinogen. PofibC1, C2, C3 and H degraded preferentially A alpha-chains while PofibS cleaved concomitantly A alpha and B beta-chains of fibrinogen. None of these enzymes cleaved the gamma-chain of fibrinogen. When correlated with the thrombin delay time, the most active was PofibS, while PofibH and PofibC1 showed almost no activity. The proteinases also differed in the peptide cleavage of B-chain of insulin. Philodryas olfersii venom promoted in vivo a loss of the circulant plasma fibrinogen, as was observed in experiments with rats.
Assakura
M T
MT
Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brazil.
Reichl
A P
AP
Mandelbaum
F R
FR
D000595:Amino Acid Sequence; D000818:Animals; D002364:Caseins / Q000378:metabolism; D017812:Colubridae; D004591:Electrophoresis, Polyacrylamide Gel; D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007525:Isoelectric Focusing; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D012910:Snake Venoms / Q000201:enzymology* / Q000633:toxicity; D013918:Thrombin Time
D002364:Caseins; D007328:Insulin; D012910:Snake Venoms; D005337:Fibrin; D005340:Fibrinogen