The PI(3,5)P2 and PI(4,5)P2 Interactomes

A comprehensive analysis of the phosphoinositide interactome has been performed using analogues of PI(3,5)P2 and PI(4,5)P2 phosphatidyl phospholipids which were immobilized onto Affi-10 beads or incorporated into liposomes for use as affinity absorbents with cytosolic extracts from colonic carcinoma cell lines. Affinity/LC/MS/MS experiments allowed identification of 388 proteins/protein complexes that appeared to interact specifically with the phosphoinositide targets: a number of novel potential phosphoinositide interacting proteins have been identified.本研究针对磷酸肌醇相互作用组（phosphoinositide interactome）开展了系统性全面分析：采用PI(3,5)P₂与PI(4,5)P₂两种磷脂酰肌醇磷酸类似物，将其固定于Affi-10微球（Affi-10 beads）上，或掺入脂质体（liposomes）中作为亲和吸附剂，与结肠癌细胞系（colonic carcinoma cell lines）的胞浆提取物（cytosolic extracts）进行孵育结合。通过亲和液相色谱-串联质谱（Affinity/LC/MS/MS）实验，共鉴定出388种可与目标磷酸肌醇特异性结合的蛋白质/蛋白质复合物，其中发现了多种全新的潜在磷酸肌醇相互作用蛋白。