Kókai Endre, Tantos Agnes, Vissi Emese, Szöor Balázs, Tompa Péter, Gausz János, Alphey Luke, Friedrich Péter, Dombrádi Viktor
Department of Medical Chemistry, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, H-4032 Debrecen, Egyetem tér 1, Hungary.
Arch Biochem Biophys. 2006 Jul 1;451(1):59-67. doi: 10.1016/j.abb.2006.03.020. Epub 2006 Apr 5.
Protein phosphatase Y (PPY) is a Drosophila testis-specific enzyme of unknown function. In a yeast two-hybrid screen we identified CG15031/PPYR1 as a PPY interacting protein. The specificity of the protein-protein interaction was proven by directed two-hybrid tests. The complex formation between PPY and PPYR1 was confirmed under in vitro and in vivo conditions by plasmon resonance spectroscopy, co-immunoprecipitation, and pull down experiments. Recombinant PPYR1 expressed in Escherichia coli is a heatstable, protease sensitive, intrinsically unstructured RNA-binding protein that migrates anomalously in SDS-polyacrylamide gel electrophoresis. It can be phosphorylated by cAMP-dependent protein kinase in vitro. PPYR1 moderately inhibits PPY activity, the inhibitory potential of the protein is slightly increased by phosphorylation. We suggest that PPYR1 may function as a scaffolding protein that targets PPY to RNA and other protein partners in Drosophila melanogaster.
蛋白磷酸酶Y(PPY)是一种功能未知的果蝇睾丸特异性酶。在酵母双杂交筛选中,我们鉴定出CG15031/PPYR1为与PPY相互作用的蛋白。通过直接双杂交试验证明了蛋白质-蛋白质相互作用的特异性。通过等离子体共振光谱、免疫共沉淀和下拉实验在体外和体内条件下证实了PPY和PPYR1之间的复合物形成。在大肠杆菌中表达的重组PPYR1是一种热稳定、蛋白酶敏感、内在无序的RNA结合蛋白,在SDS-聚丙烯酰胺凝胶电泳中迁移异常。它在体外可被cAMP依赖性蛋白激酶磷酸化。PPYR1适度抑制PPY活性,磷酸化会使该蛋白的抑制潜力略有增加。我们认为PPYR1可能作为一种支架蛋白,将PPY靶向黑腹果蝇中的RNA和其他蛋白质伴侣。