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Purification and Characterization of the Disintegrin From Gloydius Brevicaudus Venom

Author: ChenYan
Tutor: XuYunLu;LiuGuangFen
School: Fujian Medical
Course: Pharmacology
Keywords: Short-tailed snake venom Disintegrin Platelet aggregation Human umbilical vein endothelial cell line Chorioallantoic membrane Angiogenesis
CLC: R99
Type: Master's thesis
Year: 2005
Downloads: 75
Quote: 0
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Abstract


Disintegrin class contains fine - Gly - Asp (RGD) sequence or Lai - Gan - aspartic acid (KGD) sequence small molecule peptide can inhibit rely on this recognition sites containing RGD or KGD integrin with its ligand , thereby inhibiting platelet aggregation, such as angiogenesis and tumor metastasis . In this paper, short-tailed venomous snake venom disintegrin (Gloydius brevicaudus) production from Jiangsu and Zhejiang , and the determination of some of its physicochemical properties and biological activity , to provide experimental evidence for the development and utilization . Purification and some physical and chemical properties of the disintegrin determination application Superdex 75 molecular sieve chromatography , was isolated from Jiangsu and Zhejiang Agkistrodon venom 6 protein peaks ; the protein peak IV can inhibit adenosine diphosphate (ADP) -induced platelet aggregation and a molecular weight of less than 10kDa, preliminary identification as disintegrin component . The component and then by Sephadex G-25 gel filtration , the DEAE Sepharose Fast Flow ion-exchange chromatography on Lichrospher C18 reverse phase chromatography to give a solution from the prime -pure product , tentatively named as GBV Ⅳ 4 . Gbv Ⅳ 4 in gel electrophoresis , SDS-PAGE (Tris-Tricine system) , showed a single protein band , its molecular weight was calculated to be 8746 daltons by gel imaging system software ; discoid electrophoresis using polyacrylamide gel determination of the isoelectric point was 6.3. Automatic potentiometric titration phospholipase A 2 the vitality and Rick methods protease activity , the results are zero the Gbv IV on platelet aggregation using the classic method of Born measured Gbv Ⅳ 4 ADP , thrombin induced platelet aggregation . The results show the Gbv Ⅳ ADP and thrombin-induced platelet aggregation was inhibited , and showed a clear dose-effect relationship , the median inhibitory concentration ( IC 50 ) 0.22μg/ml and 1.327μg / ml. The 3 . Gbv IV in human gastric cancer cell line (SGC-7901) the growth impact of using the SRB Act IV of gastric cancer cell line SGC-7901 inhibits the proliferation observed Gbv . The results found Gbv IV can inhibit growth of SGC-7901 cells , and with the increase in drug concentration , inhibition also enhanced inhibition enhanced obvious , but increases with time . Gbv Ⅳ 4 for 24h SGC-7901 cells IC 50 to 71.68μg/ml .

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