网格蛋白调节PIPKIγ661与AP-2衔接蛋白β2附属物的结合。
Clathrin regulates the association of PIPKIgamma661 with the AP-2 adaptor beta2 appendage.
作者信息
Thieman James R, Mishra Sanjay K, Ling Kun, Doray Balraj, Anderson Richard A, Traub Linton M
机构信息
Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261.
Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905.
出版信息
J Biol Chem. 2009 May 15;284(20):13924-13939. doi: 10.1074/jbc.M901017200. Epub 2009 Mar 14.
The AP-2 clathrin adaptor differs fundamentally from the related AP-1, AP-3, and AP-4 sorting complexes because membrane deposition does not depend directly on an Arf family GTPase. Instead phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) appears to act as the principal compartmental cue for AP-2 placement at the plasma membrane as well as for the docking of numerous other important clathrin coat components at the nascent bud site. This PtdIns(4,5)P(2) dependence makes type I phosphatidylinositol 4-phosphate 5-kinases (PIPKIs) lynchpin enzymes in the assembly of clathrin-coated structures at the cell surface. PIPKIgamma is the chief 5-kinase at nerve terminals, and here we show that the 26-amino acid, alternatively spliced C terminus of PIPKIgamma661 is an intrinsically unstructured polypeptide that binds directly to the sandwich subdomain of the AP-2 beta2 subunit appendage. An aromatic side chain-based, extended interaction motif that also includes the two bulky C-terminal residues of the short PIPKIgamma635 variant is necessary for beta2 appendage engagement. The clathrin heavy chain accesses the same contact surface on the AP-2 beta2 appendage, but because of additional clathrin binding sites located within the unstructured hinge segment of the beta2 subunit, clathrin binds the beta2 chain with a higher apparent affinity than PIPKIgamma661. A clathrin-regulated interaction with AP-2 could allow PIPKIgamma661 to be strategically positioned for regional PtdIns(4,5)P(2) generation during clathrin-coated vesicle assembly at the synapse.
衔接蛋白AP-2与相关的AP-1、AP-3和AP-4分选复合物存在根本差异,因为膜沉积并不直接依赖于Arf家族GTP酶。相反,磷脂酰肌醇4,5-二磷酸(PtdIns(4,5)P(2))似乎是AP-2定位于质膜以及众多其他重要网格蛋白包被成分对接新生芽位点的主要区域线索。这种对PtdIns(4,5)P(2)的依赖性使得I型磷脂酰肌醇4-磷酸5-激酶(PIPKIs)成为细胞表面网格蛋白包被结构组装中的关键酶。PIPKIγ是神经末梢的主要5-激酶,我们在此表明,PIPKIγ661的26个氨基酸的可变剪接C末端是一种内在无序的多肽,它直接结合到AP-2β2亚基附属物的三明治亚结构域。一个基于芳香侧链的延伸相互作用基序,其中还包括短PIPKIγ635变体的两个大的C末端残基,对于β2附属物的结合是必需的。网格蛋白重链可接触到AP-2β2附属物上的相同接触表面,但由于β2亚基无序铰链段内存在额外的网格蛋白结合位点,网格蛋白与β2链的结合亲和力比PIPKIγ661更高。与AP-2的网格蛋白调节相互作用可能使PIPKIγ661在突触处网格蛋白包被囊泡组装过程中能够策略性地定位以进行局部PtdIns(4,5)P(2)生成。