一种新去整合素Adinbitor的表达、序列分析及其对C6神经胶质瘤细胞凋亡的诱导

Adinbitor为一通过基因工程方法获得的来自于中国白眉蝮蛇毒腺的去整合素家族的新成员 .从蝮蛇毒腺中提取总RNA进行RT -PCR扩增 ,获得了 2 19bp的白眉蝮蛇去整合素基因 .cDNA测序及由此推导的氨基酸序列结果显示 ,Adinbitor为含有 73个氨基酸的多肽 ,其中包括 12个半胱氨酸 ,并具有去整合素的特征模序 -RGD .对此去整合素基因进行克隆、转化与诱导后 ,得到了该蛋白的可溶性高效表达 .经组氨酸亲和层析纯化 ,获得了分子量Mr为 9×10 3 的均质融合蛋白 ,并将其命名为adinbitor.序列分析表明 ,adinbitor与已发表于GENBANK上的整合素序列有很高的同源性 ,其中同源性最高的是南韩的saxatilin ,二者的蛋白质同源性高达 91.78% .活性测定结果表明 ,Adinbitor能有效地诱导神经胶质瘤细胞C6的细胞凋亡 ,有希望成为一种抗癌新药 .图 5参 15

EXPRESSION AND SEQUENCE ANALYSES OF A NOVEL DISINTEGRIN ADINBITOR AND ITS APOPTOSIS INDUCING EFFECT ON C6 GLIOMA CELLS

Adinbitor is a recombinant protein of disintegrin from Agkistrodon halys brevicaudus stejneger. In this study, total RNA was extracted from venom gland and was reversely transcribed to cDNA which is 219 bp long. Sequencing of cDNA and deduced amino sequence indicates that the adinbitor is a polypeptide composed of 73 amino acids residues including 12 cysteines, as well as the motif- RGD. After cloning, adinbitor was expressed in E.coli and was purified by using the His-Bind Column Chromatography under native condition. The soluble homogeneous protein was obtained. Sequence analyses indicated that adinbitor′s amino acids sequence was 91.78% homology with saxatilin. Some results showed that adinbitor could induce apoptosis of C6 glioma cells. Fig 5, Ref 15