A novel P-I class metalloproteinase with broad substrate-cleaving activity, agkislysin, from Agkistrodon acutus venom.
The venom of Viperdae is a rich source of metalloproteinases, which have potential clinical applications for lowering plasma fibrinogen or dissolving thrombi. Recently, we purified a novel proteinase from Formosan Agkistrodon acutus venom using DEAE-Sephadex A-50 and Mono-Q HR 5/5 column chromatography. The purified getatinolytic component, agkislysin, is a 22kDa-monomeric protein without Asn-linked sugar. Functional characterization showed that agkislysin possessed both fibronectin- and type IV collagen-cleaving activities. In addition, agkislysin preferentially cleaved the Aalpha chain of fibrinogen, followed by the Bbeta chain, while the gamma chain was finally affected. Furthermore, agkislysin was also capable of cleaving prothrombin into various fragments, as well as suppressing ristocetin-induced platelet aggregation by hydrolyzing von Willebrand factor. However, the proteolytic activity of agkislysin was slightly enhanced by divalent metal ions and completely inhibited by chelating agents, but not serine proteinase inhibitor. These findings suggest that agkislysin is a P-I class metalloproteinase with unique biological properties.
Wang
Wen-Jeng
WJ
Chang-Gung Institute of Technology, Kwei-Shan,Tao-Yuan 33303, Taiwan, ROC. wjwang@mail.cgit.edu.tw <wjwang@mail.cgit.edu.tw>
Shih
Chun-Ho
CH
Huang
Tur-Fu
TF
D017836:Agkistrodon / Q000378:metabolism*; D000818:Animals; D002364:Caseins / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000302:isolation & purification* / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D045726:Metalloproteases / Q000302:isolation & purification* / Q000378:metabolism*; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification / Q000378:metabolism; D011516:Prothrombin / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology
C031442:Agkistrodon venoms; D002364:Caseins; D024141:Collagen Type IV; D003435:Crotalid Venoms; D010975:Platelet Aggregation Inhibitors; C026018:azocasein; D012310:Ristocetin; D011516:Prothrombin; D005340:Fibrinogen; D045726:Metalloproteases; C494429:agkislysin, Agkistrodon acutus