Zhou Xing-Ding, Ding Chen-Hui, Tai Hong, Jin Yang, Chen Run-Qiang, Lu Qiu-Min, Wang Wan-Yu, Xiong Yu-Liang
Department of Animal Toxinology, Kunming Institute of Zoology, The Chinese Academy of Sciences, No. 32 Jiaochangdonglu, Kunming 650223, Yunnan, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2004 Sep;139(1):117-22. doi: 10.1016/j.cbpc.2004.06.012.
A novel disintegrin, jerdonatin, was purified to homogeneity from Trimeresurus jerdonii venom by gel filtration and reversed-phase high-pressure liquid chromatography. We isolated the cDNA encoding jerdonatin from the snake venom gland. Jerdonatin cDNA precursor encoded pre-peptide, metalloprotease and disintegrin domain. Jerdonatin is composed of 72 amino acid residues including 12 cysteines and the tripeptide sequence Arg-Gly-Asp (RGD), a well-known characteristic of the disintegrin family. Molecular mass of jerdonatin was determined to be 8011 Da by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Jerdonatin inhibited ADP- and collagen-induced human platelet aggregation with IC50 of 123 and 135 nM, respectively. We also investigated the effect of jerdonatin on the binding of B6D2F1 hybrid mice spermatozoa to mice zona-free eggs and their subsequent fusion. Jerdonatin significantly inhibited sperm-egg binding in a concentration-dependent manner, but had no effect on the fusion of sperm-egg. These results indicate that integrins on the egg play a role in mammalian fertilization.
通过凝胶过滤和反相高效液相色谱法,从竹叶青蛇毒中纯化出一种新型去整合素——杰氏去整合素,使其达到同质状态。我们从蛇毒腺中分离出编码杰氏去整合素的cDNA。杰氏去整合素cDNA前体编码前肽、金属蛋白酶和去整合素结构域。杰氏去整合素由72个氨基酸残基组成,包括12个半胱氨酸以及三肽序列精氨酸-甘氨酸-天冬氨酸(RGD),这是去整合素家族的一个众所周知的特征。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)测定,杰氏去整合素的分子量为8011道尔顿。杰氏去整合素抑制二磷酸腺苷(ADP)和胶原蛋白诱导的人血小板聚集,其半数抑制浓度(IC50)分别为123和135纳摩尔。我们还研究了杰氏去整合素对B6D2F1杂交小鼠精子与去透明带小鼠卵子结合及其随后融合过程的影响。杰氏去整合素以浓度依赖的方式显著抑制精卵结合,但对精卵融合没有影响。这些结果表明,卵子上的整合素在哺乳动物受精过程中发挥作用。