%0 Journal Article
%T Expression,Purification of PUMA-BH3 Death Domain Peptide in E.coli and Identification of Its Pro-apoptotic Activity<br>PUMA-BH3结构域短肽的原核表达、纯化及促凋亡活性鉴定
%A ZHANG Yu-wen
%A LIU Xing-han
%A LIN Hui-min
%A LI Ji-hong
%A MA Hong-xing
%A LIU Yuan-li
%A <br>张宇雯
%A 刘兴汉
%A 林慧敏
%A 李冀红
%A 马洪星
%A 刘远莉
%J 中国生物工程杂志
%D 2007
%I 
%X The Bcl-2 family of proteins play a central role in the control of apoptosis, a fundamental process for both human health and disease, by mitochondrial pathway. PUMA(p53 up-regulated modulator of apoptosis protein) is one of BH3-only members of Bcl-2 family , its function is to promote cell apoptosis. To obtain BH3 death domain peptide of PUMA and detect its biological activity, the synthesized double-stranded oligomeric nucleotide encoding PUMA-BH3 peptide was cloned into expression vector pTYB2,thus generating a construct of pTYB2-PUMA-BH3 which expressed PUMA-BH3-intein-chitin binding domain fusion protein. Then the recombinant plasmid was transformed into E.coli BL-21 (DE3) and fusion protein was expressed under induction by IPTG. The soluble PUMA-BH3 peptide was purified from chitin affinity chromatography by DTT reduction. Through measuring mitochondria viability(MTT),mitochondria permeability transition(MPT) and the translocation of cytochrome c(Cyt c ) assayed by western blotting, the biological pro-apoptotic activity of PUMA-BH3 peptide was studied. The PUMA-BH3 peptide has the effects on decreasing the mitochondria viability remarkably , inducing mitochondrial swelling and promoting Cyt c releasing from isolated mitochodria . Mitochondrial swelling and the release of Cyt c induced by PUMA-BH3 peptide concerned with the opening of MPT,which can be improved by cyclosporine A(CsA).These results indicated that recombinant PUMA-BH3 peptide might possess pro-apoptosis activity and paved a reasonable way for the study of new apoptosis regulators.
%K BH3 death domain peptide of PUMA Recombinant expression Mitochondria Pro-apoptotic activity<br>PUMA-BH3短肽
%K 重组表达
%K 线粒体
%K 促凋亡活性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=DF315A43E6E31CB1&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=DF92D298D3FF1E6E&sid=DB817633AA4F79B9&eid=9971A5E270697F23&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=13