Purification and properties of the thrombin-like enzyme from the venom of Lachesis muta muta.
1. The coagulating enzyme of the Lachesis muta muta venom was purified to homogeneity by a combination of a gel filtration in Sephadex G-100 and affinity chromatography on agarose-agmatine resin. 2. Several forms of the enzyme were prepared by isoelectric focusing with pIs ranging from 3.1 to 5.0; the asialoenzyme focused as a narrow band at pH 8.7. SDS-PAGE analysis of the purified enzyme revealed a single broad band with apparent Mr of 41-47 kDa. 3. The enzyme cleaves only fibrinopeptide A from fibrinogen; it does not activate factor XIII and is devoid of kallikrein-like activity. 4. Kinetic properties of the enzyme were determined for representative synthetic chromogenic substrates and inhibitors.
Silveira
A M
AM
Departamento de Bioquímica e Immunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Brasil.
Magalhães
A
A
Diniz
C R
CR
de Oliveira
E B
EB
D000818:Animals; D001777:Blood Coagulation; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography / Q000379:methods; D003435:Crotalid Venoms / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods; D005340:Fibrinogen / Q000378:metabolism; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism
D003435:Crotalid Venoms; D005340:Fibrinogen; D013917:Thrombin