Purification and characterization of a non-hemorrhagic metalloprotease from Agkistrodon halys brevicaudus venom.
A non-hemorrhagic metalloprotease (protease L4) was purified from the venom of Chinese Mamushi (Agkistrodon halys brevicaudus) by gel filtration and anion-exchange chromatography. Protease L4 has the molecular weight of 22,000 and its optimum pH was 8.5. The protein was stable in the pH range of 5-9 and below 40 degrees C. The proteolytic activity was inhibited by metal-chelating agents and some metal ions. Calcium ion activated the activity dose-dependently, but had only a minor effect on the thermal and pH stability. L4 showed fibrinogenase activity, hydrolyzing only the A alpha chain of fibrinogen. The protease cleaved preferentially at the N-terminal of Leu and His residues of some peptides.
Fujimura
S
S
Department of Chemistry, Faculty of Science, Fukuoka University, Japan.
Rikimaru
T
T
Baba
S
S
Hori
J
J
Hao
X Q
XQ
Terada
S
S
Kimoto
E
E
D017836:Agkistrodon; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002412:Cations / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D002845:Chromatography; D004795:Enzyme Stability; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006470:Hemorrhage; D006868:Hydrolysis; D008666:Metalloendopeptidases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D014757:Viper Venoms / Q000201:enzymology*
D000596:Amino Acids; D002412:Cations; D002614:Chelating Agents; D014757:Viper Venoms; D005337:Fibrin; D005340:Fibrinogen; D008666:Metalloendopeptidases