Identification of acceptor proteins and their modification sites of polyADP-ribosylation and biological function

已知多聚ADP核糖化的生物学功能之一与DNA碱基切除修复有关。然而，其他功能仍不清楚。为了通过鉴定大多数未知的体内多聚ADP-核糖化受体蛋白来阐明其功能，我们选择了缺失多聚ADP-核糖降解酶基因的黑腹果蝇突变体。这种突变的果蝇在高温下会出现皱纹，并表现出神经系统疾病。因此，这可能会成为人类神经系统疾病的模型。利用固定化抗多聚ADP-核糖抗体亲和层析柱、碱性处理、双向凝胶电泳法和质谱法，我们在体内鉴定了乙醇脱氢酶等几种蛋白质为多ADP-核糖化的候选蛋白。

One of the biological functions of polyADP-ribosylation has been known to be involved in DNA base excision repair. However the other functions are still not known yet. To clarify the functions through identifying the in vivo acceptor proteins of polyADP-ribosylation that is mostly unknown, we used Drosophila melanogaster mutant that had deleted poly(ADP-ribose) degradation enzyme gene, (poly(ADP-ribose) glycohydrolase gene. This mutant Drosophila could eclose under elevated temperature and shows neurological disease. Thus this might serve as a model for human neurological diseases. Using affinity column of immobilized antibody against poly(ADP-ribose), alkaline treatment, 2D gel electrophoresis and mass spectroscopy, we identified alcohol dehydrogenase and several other proteins as candidates of polyADP-ribosylation in vivo.