Molecular cloning and expression of catrocollastatin, a snake-venom protein from Crotalus atrox (western diamondback rattlesnake) which inhibits platelet adhesion to collagen.
A 50 kDa protein that inhibits platelet adhesion to collagen has been isolated from snake venom of Crotalus atrox (western diamondback rattlesnake) and has been named 'catrocollastatin'. The cDNA cloning of catrocollastatin has been accomplished. A full-length cDNA of 2310 bp with an open reading frame between nucleotides 51 and 1880 was obtained. The deduced amino acid sequence consists of 609 amino acids. The cDNA-predicted amino acid sequence is highly similar to that of haemorrhagic metalloproteinase jararhagin from Bothrops jararaca venom, HR1B from Trimeresurus flavoviridis, Ht-e from C. atrox and trigramin from T. gramineus. Like jararhagin and HR1B, catrocollastatin is a multidomain molecule composed of an N-terminal domain, a metalloproteinase domain, a disintegrin-like domain and a cysteine-rich C-terminal domain. In the disintegrin-like domain, the frequently seen RGD (Arg-Gly-Asp) sequence is replaced by SECD (Ser-Glu-Cys-Asp). This cDNA was expressed in Spodoptera frugiperda (fall armyworm) (Sf9) insect cells using a baculovirus expression system. Like native catrocollastatin, the expressed protein is capable of selectively blocking collagen-induced platelet aggregation. This is the first full-length clone of a high-molecular-mass haemorrhagin to be expressed.
Zhou
Q
Q
Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Smith
J B
JB
Grossman
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D003094:Collagen / Q000378:metabolism*; D003435:Crotalid Venoms / Q000737:chemistry*; D004247:DNA; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D008969:Molecular Sequence Data; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera
D003435:Crotalid Venoms; D010975:Platelet Aggregation Inhibitors; D003094:Collagen; D004247:DNA; D008666:Metalloendopeptidases; C093237:catrocollastatin