Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices
Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices
复制标题
DOI：
10.1016/s1097-2765(02)00434-3
复制
发表时间：
2002-01-01
期刊：
影响因子：
16
通讯作者：
Hawkins, PT
中科院分区：
文献类型：
--
作者：
Krugmann, S;Anderson, KE;Hawkins, PT
我们表明，携带天然磷酸肌醇同系物的基质可用于捕获和展示细胞和组织提取物中的多种磷酸肌醇结合蛋白。我们展示了通过这种方法分离的 20 多种蛋白质的质谱鉴定，大部分来自白细胞提取物：它们包括具有已确定的磷酸肌醇结合域的已知和新型蛋白质，以及具有令人惊讶和不寻常的磷酸肌醇结合特性的已知蛋白质。 ARAP3 是一种新型 PtdIns(3,4,5)P-3 结合蛋白，具有不寻常的结构域结构，包括五个预测的 PH 结构域。我们证明它是体外和体内特异性 PtdIns(3,4,5)P-3/PtdIns(3,4)P-2 刺激的 Arf6 GAP，并且其 Arf GAP 和 Rho GAP 结构域协同介导细胞骨架和细胞形状中 PI3K 依赖性重排。
We show that matrices carrying the tethered homologs of natural phosphoinositides can be used to capture and display multiple phosphoinositide binding proteins in cell and tissue extracts. We present the mass spectrometric identification of over 20 proteins isolated by this method, mostly from leukocyte extracts: they include known and novel proteins with established phosphoinositide binding domains and also known proteins with surprising and unusual phosphoinositide binding properties. One of the novel PtdIns(3,4,5)P-3 binding proteins, ARAP3, has an unusual domain structure, including five predicted PH domains. We show that it is a specific PtdIns(3,4,5)P-3/PtdIns(3,4)P-2-stimulated Arf6 GAP both in vitro and in vivo, and both its Arf GAP and Rho GAP domains cooperate in mediating PI3K-dependent rearrangements in the cell cytoskeleton and cell shape.