Finniss Susan, Movsisyan Ashley, Billecke Christine, Schmidt Mirko, Randazzo Lisa, Chen Baihua, Bögler Oliver
William and Karen Davidson Laboratory of Brain Tumor Biology, Hermelin Brain Tumor Center, Department of Neurosurgery, Henry Ford Hospital, 2799 West Grand Blvd., Detroit, MI 48202, USA.
Biochem Biophys Res Commun. 2004 Dec 3;325(1):174-82. doi: 10.1016/j.bbrc.2004.10.007.
SETA/CIN85/Ruk is a multifunctional adaptor protein involved in signal transduction and attenuation downstream of receptor tyrosine kinases. It has a modular structure, and various isoforms that combine different protein-protein interaction domains have been proposed based on cDNA analysis. As a first step towards understanding SETA/CIN85/Ruk isoforms at the protein level, we have characterized 5 monoclonal antibodies against this protein. Three of these were used to study lysates fractionated on a pH gradient, leading to the identification of various SETA/CIN85/Ruk proteins on the basis of pI and apparent molecular weight. While good correspondence with proteins predicted from cDNA analysis was found for two isoforms, in most cases it was not possible to make an unequivocal assignment. We conclude that additional splice variants remain to be described, and that a deeper understanding of SETA/CIN85/Ruk post-translational processing and modification is necessary to gain further understanding of this complex gene product.
SETA/CIN85/Ruk是一种多功能衔接蛋白，参与受体酪氨酸激酶下游的信号转导和信号衰减。它具有模块化结构，基于cDNA分析提出了结合不同蛋白质-蛋白质相互作用结构域的各种异构体。作为在蛋白质水平上理解SETA/CIN85/Ruk异构体的第一步，我们鉴定了5种针对该蛋白的单克隆抗体。其中3种用于研究在pH梯度上分级分离的裂解物，从而根据等电点和表观分子量鉴定出各种SETA/CIN85/Ruk蛋白。虽然发现两种异构体与cDNA分析预测的蛋白质有很好的对应关系，但在大多数情况下，无法做出明确的归属。我们得出结论，仍有待描述其他剪接变体，并且有必要更深入地了解SETA/CIN85/Ruk的翻译后加工和修饰，以进一步了解这种复杂的基因产物。
