Abstract
Adinbitor was cloned from Agkistrodon halys brevicaudus stejneger and characterized as a novel disintegrin. In this study, total RNA was extracted from venom gland and used in RT-PCR to generate a cDNA which is 219 bp long. The sequence encoded a polypeptide composed of 73 amino acids, including 12 cysteines, an RGD motif, and the signature motif of disintergrin. Recombinant Adinbitor (rAdinbitor) was expressed in E. coli and purified by using the His. Bind affinity chromatography. The IC(50) for inhibiting human platelet aggregation and bFGF-induced proliferation of ECV304 cells was 6 microM and 0.89 microM respectively. Furthermore, Adinbitor significantly inhibited angiogenesis both in vivo and in vitro. Taken together, these results suggested that Adinbitor had typical functions of disintegrins.
MeSH terms
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Agkistrodon
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Amino Acid Motifs
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Amino Acid Sequence
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Animals
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Base Sequence
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Cell Proliferation
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Cell Survival
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Chick Embryo
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Chromatography
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Cloning, Molecular
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Collagen / pharmacology
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Crotalid Venoms / chemistry*
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Crotalid Venoms / genetics*
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Crotalid Venoms / metabolism
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DNA, Complementary / metabolism
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Disintegrins / chemistry*
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Disintegrins / genetics*
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Dose-Response Relationship, Drug
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Drug Combinations
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Electrophoresis, Polyacrylamide Gel
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Fibroblast Growth Factor 2 / metabolism
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Genetic Vectors
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Inhibitory Concentration 50
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Laminin / pharmacology
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Methylcellulose / chemistry
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Molecular Sequence Data
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Neovascularization, Pathologic
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Oligopeptides / chemistry
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Platelet Aggregation
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Proteoglycans / pharmacology
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RNA / chemistry
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Recombinant Proteins / chemistry
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Reverse Transcriptase Polymerase Chain Reaction
Substances
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Agkistrodon venoms
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Crotalid Venoms
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DNA, Complementary
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Disintegrins
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Drug Combinations
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Laminin
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Oligopeptides
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Proteoglycans
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Recombinant Proteins
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adinbitor protein, Agkistrodon halys brevicaudus stejneger
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Fibroblast Growth Factor 2
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matrigel
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RNA
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arginyl-glycyl-aspartic acid
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Methylcellulose
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Collagen