Bioassay-directed purification of an acidic phospholipase A(2) from Agkistrodon halys pallas venom.
Relying on ex vivo and in vitro platelet anti-aggregation assays, a tail bleeding time assay, and an anti-thrombotic assay, we have purified the fraction of venom from Agkistrodon halys pallas which, in all of these assays, is the most active. There were two major steps in the purification: gel filtration chromatography on Sephadex G-100, and ion exchange chromatography on DEAE Sephadex A-50. Sequencing of the most active fraction by mass spectrometry revealed that it is a known acidic phospholipase A(2). Prior expectations by others about the in vivo anti-thrombotic activity of this enzyme are confirmed.
Wang
Yuwei
Y
College of Pharmaceutical Sciences, Capital Medical University, Beijing 100069, PR China.
Cui
Guohui
G
Zhao
Ming
M
Yang
Jian
J
Wang
Chao
C
Giese
Roger W
RW
Peng
Shiqi
S
D017836:Agkistrodon; D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification* / Q000494:pharmacology; D001681:Biological Assay; D001760:Bleeding Time; D005591:Chemical Fractionation; D002845:Chromatography; D003435:Crotalid Venoms / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010449:Peptide Mapping; D054467:Phospholipases A2 / Q000302:isolation & purification* / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D013927:Thrombosis / Q000188:drug therapy
D000925:Anticoagulants; D003435:Crotalid Venoms; D054467:Phospholipases A2