Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin-mediated endocytosis.
Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin-mediated endocytosis.
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DOI：
10.1083/jcb.200108123
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发表时间：
2002-03-04
期刊：
影响因子：
--
通讯作者：
Schmid SL
中科院分区：
文献类型：
--
作者：
Conner SD;Schmid SL
已知AP 2复合物的μ2亚基在体外被共纯化激酶磷酸化，并且最近已经证明转铁蛋白内吞作用需要μ2磷酸化（Olusanya，O.，警局安德鲁，J.R. Swedlow和E.斯迈思2001. Curr. 11：896-900）。然而，负责这种磷酸化的内源性激酶的身份是未知的。在这里，我们确定和表征的Prk/方舟家庭的丝氨酸/苏氨酸激酶，衔接子相关激酶（AAK）1的新成员。我们发现AAK 1与接头蛋白（AP）2共纯化，并在体内和体外直接结合α-adaptin的耳域。在神经元细胞中，AAK 1富集在突触前末梢，而在非神经元细胞中，它与网格蛋白和AP 2共定位在网格蛋白包被的凹坑和迁移细胞的前缘。AAK 1在体外特异性磷酸化μ亚基，内吞作用的阶段特异性测定表明，AAK 1的μ磷酸化导致AP 2刺激的转铁蛋白内化减少。总之，这些结果提供了强有力的证据，AAK 1是内源性μ2激酶，并在网格蛋白介导的内吞作用中发挥调节作用。这些结果也支持网格蛋白介导的内吞作用是由磷酸化/去磷酸化循环控制的观点。
The μ2 subunit of the AP2 complex is known to be phosphorylated in vitro by a copurifying kinase, and it has been demonstrated recently that μ2 phosphorylation is required for transferrin endocytosis (Olusanya, O., P.D. Andrews, J.R. Swedlow, and E. Smythe. 2001. Curr. Biol. 11:896–900). However, the identity of the endogenous kinase responsible for this phosphorylation is unknown. Here we identify and characterize a novel member of the Prk/Ark family of serine/threonine kinases, adaptor-associated kinase (AAK)1. We find that AAK1 copurifies with adaptor protein (AP)2 and that it directly binds the ear domain of α-adaptin in vivo and in vitro. In neuronal cells, AAK1 is enriched at presynaptic terminals, whereas in nonneuronal cells it colocalizes with clathrin and AP2 in clathrin-coated pits and at the leading edge of migrating cells. AAK1 specifically phosphorylates the μ subunit in vitro, and stage-specific assays for endocytosis show that μ phosphorylation by AAK1 results in a decrease in AP2-stimulated transferrin internalization. Together, these results provide strong evidence that AAK1 is the endogenous μ2 kinase and plays a regulatory role in clathrin-mediated endocytosis. These results also lend support to the idea that clathrin-mediated endocytosis is controlled by cycles of phosphorylation/desphosphorylation.
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DOI：
10.1083/jcb.144.6.1203
发表时间：
1999-03-22
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cope MJ;Yang S;Shang C;Drubin DG
通讯作者：
Drubin DG
影响因子：
4.8
作者：
Bauerfeind, R;Takei, K;De Camilli, P
通讯作者：
De Camilli, P
影响因子：
11.4
作者：
AHLE, S;MANN, A;UNGEWICKELL, E
通讯作者：
UNGEWICKELL, E
影响因子：
56.9
作者：
LIU, JP;SIM, ATR;ROBINSON, PJ
通讯作者：
ROBINSON, PJ
影响因子：
4.1
作者：
PAULOIN, A;THURIEAU, C
通讯作者：
THURIEAU, C