A fibrinogen-clotting serine proteinase from Cerastes cerastes (horned viper) venom with arginine-esterase and amidase activities. Purification, characterization and kinetic parameter determination.
An enzyme displaying proteolytic activity toward the natural substrate casein as well as clotting activity on fibrinogen was purified to homogeneity from Cerastes cerastes (horned viper) venom and characterized. The enzyme is constituted of two identical subunits of mol. wt 48,500 as determined by SDS-polyacrylamide gel electrophoresis, and has an isoelectric point of 3.75. N-terminal sequencing up to the 33rd residue evidenced a high homology with other snake venom proteinases. The proteinase is of serine-type as indicated by high sensitivity to DFP and shows both arginine-ester hydrolase and amidase activities on synthetic substrates. Both specific activities were 30-fold higher than the respective activities found in the crude venom. The Km value determined for arginine-containing substrate BAEE was 3.0 x 10(-4) M and the Km for chromogenic substrate CBS 34-47 0.65 x 10(-4) M. The Vm/Km ratio, however, was two-fold higher for BAEE than for CBS 34-47; the arginine-esterase activity of this enzyme is thus slightly higher than its amidase activity.
Laraba-Djebari
F
F
Centre National de la Recherche Scientifique, Laboratoire de Biochimie, Faculté de Médecine, Marseille, France.
Martin-Eauclaire
M F
MF
Marchot
P
P
D000581:Amidohydrolases / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D002364:Caseins / Q000378:metabolism; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000187:drug effects*; D007525:Isoelectric Focusing; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D014757:Viper Venoms / Q000201:enzymology / Q000494:pharmacology*
D002364:Caseins; D014757:Viper Venoms; D007531:Isoflurophate; D005340:Fibrinogen; C019724:arginine esterase; D002265:Carboxylic Ester Hydrolases; D012697:Serine Endopeptidases; D000581:Amidohydrolases; C019665:amidase