VGD and MLD-motifs containing heterodimeric disintegrin viplebedin-2 from Vipera lebetina snake venom. Purification and cDNA cloning.
We have previously demonstrated that the fibrinolytic enzyme lebetase is synthesized with disintegrin-like domain that is cleaved posttranslationally (Siigur et al., 1996). Now we isolated a heterodimeric disintegrin viplebedin-2 containing this disintegrin-like part from Vipera lebetina venom using size-exclusion chromatography on Sephadex G-100 sf and HPLC on C18 column. The molecular masses of viplebedin-2 and tryptic peptides from both chains of viplebedin-2 were determined by MALDI-TOF mass spectrometry. Using cDNA library of the venom gland of a single V. lebetina turanica snake the viplebedin-2 coding cDNAs were cloned and sequenced. Viplebedin-2 chains are synthesized from two different genes. One chain, containing VGD sequence in disintegrin loop, is synthesized as a disintegrin-like part of the PII-type metalloprotease, lebetase. The other chain, containing MLD sequence in disintegrin loop, is synthesized from the gene without metalloproteinase domain. Two polyadenylation signal sequences have been found in MLD sequence coding chain precursor cDNAs. Viplebedin-2 dose-dependently inhibited adhesion of platelets to immobilized collagen and inhibited collagen-induced platelet aggregation.
Vija
Heiki
H
National Institute of Chemical Physics and Biophysics, Akadeemia tee 23,Tallinn 12618, Estonia.
Samel
Mari
M
Siigur
Ene
E
Aaspõllu
Anu
A
Tõnismägi
Külli
K
Trummal
Katrin
K
Subbi
Juhan
J
Siigur
Jüri
J
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D019483:Disintegrins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014757:Viper Venoms / Q000378:metabolism / Q000494:pharmacology; D017819:Viperidae / Q000378:metabolism*
D018076:DNA, Complementary; D019483:Disintegrins; D010975:Platelet Aggregation Inhibitors; D014757:Viper Venoms