Fan Jing, Zhang Yu-Qing, Li Ping, Tong Chang, Tan Li, Zhu Yun-Song
Department of Molecular Genetics, Shanghai Medical School of Fudan University, Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai 200032, China.
Acta Biochim Biophys Sin (Shanghai). 2004 Sep;36(9):623-8. doi: 10.1093/abbs/36.9.623.
The plasminogen activator inhibitor type-2 (PAI-2) dependent apoptosis protection is due to the 33 amino acids fragment located between helix C and D of PAI-2, this fragment may interact with some unknown intracellular proteins. In this study we used the fragment between helix C and D of PAI-2 as a bait to perform a yeast two-hybrid screen using a cDNA library constructed with HeLa cells during apoptosis, and retrieved a clone encoding 94 amino acid residues of C-terminus of pre-mRNA processing factor 8 (PRPF8). Co-immunoprecipitation experiments confirmed that PAI-2 could interact with PRPF8 in vivo. PAI-2 could bind PRPF8 C-terminal in both the inside and outside of nuclear. These results suggested that the interaction between these two proteins might not be involved in the apoptosis process.
纤溶酶原激活物抑制剂-2(PAI-2)依赖的细胞凋亡保护作用归因于PAI-2的C螺旋和D螺旋之间的33个氨基酸片段,该片段可能与一些未知的细胞内蛋白质相互作用。在本研究中,我们使用PAI-2的C螺旋和D螺旋之间的片段作为诱饵,利用凋亡过程中HeLa细胞构建的cDNA文库进行酵母双杂交筛选,获得了一个编码前体mRNA加工因子8(PRPF8)C末端94个氨基酸残基的克隆。免疫共沉淀实验证实PAI-2在体内可与PRPF8相互作用。PAI-2在细胞核内外均可结合PRPF8的C末端。这些结果表明这两种蛋白质之间的相互作用可能不参与细胞凋亡过程。