Purification and characterization of a fibrinogenase from Vipera lebetina (desert adder) venom.
A fibrinogenase from Vipera lebetina venom was isolated by gel filtration in a Superose 12 column prep grade HR 16/50 and by ion-exchange in a Mono Q HR 5/5 column. The purified enzyme, which was obtained with a yield of 8 mg from 60 mg of crude venom, is a glycoprotein having an isoelectric point of 5.9 +/- 0.1 and a mol. wt of 26,000 +/- 1000 as estimated by SDS-PAGE. The biochemical characterization of the enzyme revealed that it hydrolyzes readily the B beta chain of fibrinogen and the A alpha chain as well as fibrin and casein. Over a pH range from 4 to 11 the enzyme was not inactivated by a 20 min treatment at 90 degrees C. The isolated fibrinogenase is inhibited by ethylenediamine tetraacetic acid, dithiothreitol and L-cysteine but not by phenylmethylsulfonyl fluoride. On the other hand, it is activated by Ca2+ and Mg2+. Purified fibrinogenase up to a dose of 100 micrograms/mouse shows no toxicity and has no hemorrhagic activity.
Institut Pasteur de Tunis, Belvedere, Tunisia.
A
A
Gasmi
M
M
Karoui
Z
Z
Benlasfar
H
H
Karoui
M
M
el Ayeb
K
K
Dellagi
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002364:Caseins / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006868:Hydrolysis; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D013917:Thrombin / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity; D014757:Viper Venoms / Q000737:chemistry* / Q000633:toxicity
D002364:Caseins; D014757:Viper Venoms; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin