Interaction of apohtosis inhibitor PI-9 with cellular factors

凋亡抑制剂 PI-9 与细胞因子的相互作用

• 批准号: 13670314
• 负责人: KANAMORI Hiroshi
• 金额: $ 1.15万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670314/
• 关键词: Proteinase inhibitor-9; Phage display; アポトーシス; 肝細胞; セリン・プロテネース; PI-9

Granzyme B (GraB) is secreted from cytotoxic T cells (CTL) and ntural killer (NK) cells and plays a key role in the apoptotic reaction of the target cells. Proteinase-9 (PI-9) is known to interact with GrB and prevent cells from apoptotic attack from CTL and NK cells. Recent investigation by others has revealed the interaction of PI-9 with other cellular molecules such as caspase. I and elastase. It has been suggested that this serine proteinase has the potential of binding to several sets of intracellular molecules and has other unknown physiological roles.We have introduced a system by which PI-9 expression levels were regulated in the presence of tetracycline using HepG2 liver cells. We observed that the upregulation of PI-9 prevent the apoptotic reaction caused by GrB. Induction of PI-9 also prevented the apoptosis by the attack of human NK cell line YT. These observations enable us to utilize this system to observe the effect of the interaction of other molecule with PI-9 on cellular physiology.We have established a system by which we can produce biotinated recombinant PI-9 by using E. Coli. We have developed a phage display system to obtain oligopeptides with high affinity binding to PI-9 protein. From a library of 12-mer random oligonucleotides, we have obtained a consensus sequence (LLADTTHHRPWT). By employing a data base search (BLAST), we have obtained several known and unknown proteins in which contain homologous amino acid sequence with the consensus sequence.These results encourage us to further investigate weatherthe interaction of the individual protein with PI-9 may have significant physiological effect.

颗粒酶B（GraB）由细胞毒性T细胞（CTL）和自然杀伤细胞（NK）分泌，在靶细胞的凋亡反应中起关键作用。蛋白酶-9（PI-9）与GrB相互作用，阻止CTL和NK细胞对细胞的凋亡攻击。其他人最近的研究揭示了PI-9与其他细胞分子如胱天蛋白酶的相互作用。我和弹性蛋白酶。有人建议，这种丝氨酸蛋白酶有可能结合到几套细胞内分子，并有其他未知的生理roles.We已经介绍了一个系统，通过该系统PI-9的表达水平调节四环素的存在下，使用HepG 2肝细胞。我们观察到PI-9的上调阻止了GrB引起的凋亡反应。PI-9的诱导也阻止了由人NK细胞系YT攻击引起的凋亡。这些结果使我们能够利用该系统来观察其他分子与PI-9相互作用对细胞生理的影响。杆菌我们已经开发了噬菌体展示系统，以获得与PI-9蛋白具有高亲和力结合的寡肽。从12-mer随机寡核苷酸文库中，我们获得了一个共有序列（LLADTTHHRPWT）。通过数据库检索（BLAST），我们获得了几个已知和未知的蛋白质，它们的氨基酸序列与PI-9的共有序列具有同源性，这些结果鼓励我们进一步研究单个蛋白质与PI-9的相互作用是否具有显著的生理效应。