Regulation of oxysterol-binding protein Golgi localization through protein kinase D-mediated phosphorylation.
Regulation of oxysterol-binding protein Golgi localization through protein kinase D-mediated phosphorylation.
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DOI：
10.1091/mbc.e10-02-0090
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发表时间：
2010-07-01
影响因子：
3.3
通讯作者：
Toker A
中科院分区：
文献类型：
--
作者：
Nhek S;Ngo M;Yang X;Ng MM;Field SJ;Asara JM;Ridgway ND;Toker A
蛋白激酶D（PKD）是高尔基体结构和功能的重要调节因子。生化证据表明，氧固醇结合蛋白OSBP作为一种新的PKD底物。磷酸化抑制OSBP高尔基体定位，损害CERT高尔基体定位，并促进高尔基体片段化。蛋白激酶D（PKD）通过调节质膜转运载体的分裂在高尔基体网络中起着关键作用。两种已知的高尔基体定位的PKD底物，PI 4-激酶IIIβ和神经酰胺转移蛋白CERT，介导PKD信号传导以分别影响囊泡向质膜的运输和鞘磷脂的合成。PKD通过与二酰基甘油相互作用在高尔基体被募集和激活，二酰基甘油是由神经酰胺合成鞘磷脂的副产物。在这里，我们确定了一个新的底物PKD在高尔基体，氧固醇结合蛋白OSBP。使用底物定向磷酸化特异性抗体，识别最佳PKD共识基序，我们表明，PKD磷酸化OSBP在Ser 240在体外和细胞中。我们进一步表明OSBP磷酸化发生在高尔基体。PKD对OSBP的磷酸化不调节二聚化、固醇结合或对PI（4）P的亲和力。相反，磷酸化响应于25-羟基胆固醇和胆固醇消耗而减弱OSBP高尔基体定位，损害CERT高尔基体定位，并促进高尔基体片段化。
Protein kinase D (PKD) is a critical regulator of Golgi structure and function. Biochemical evidence is presented that demonstrates the oxysterol-binding protein OSBP as a novel PKD substrate. Phosphorylation inhibits OSBP Golgi localization, impairs CERT Golgi localization, and promotes Golgi fragmentation. Protein kinase D (PKD) plays a critical role at the trans-Golgi network by regulating the fission of transport carriers destined for the plasma membrane. Two known Golgi-localized PKD substrates, PI4-kinase IIIβ and the ceramide transfer protein CERT, mediate PKD signaling to influence vesicle trafficking to the plasma membrane and sphingomyelin synthesis, respectively. PKD is recruited and activated at the Golgi through interaction with diacylglycerol, a pool of which is generated as a by-product of sphingomyelin synthesis from ceramide. Here we identify a novel substrate of PKD at the Golgi, the oxysterol-binding protein OSBP. Using a substrate-directed phospho-specific antibody that recognizes the optimal PKD consensus motif, we show that PKD phosphorylates OSBP at Ser240 in vitro and in cells. We further show that OSBP phosphorylation occurs at the Golgi. Phosphorylation of OSBP by PKD does not modulate dimerization, sterol binding, or affinity for PI(4)P. Instead, phosphorylation attenuates OSBP Golgi localization in response to 25-hydroxycholesterol and cholesterol depletion, impairs CERT Golgi localization, and promotes Golgi fragmentation.
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二聚体 PKD 调节膜裂变以在 TGN 处形成运输载体。
影响因子：
7.8
作者：
Bossard, Carine;Bresson, Damien;Polishchuk, Roman S.;Malhotra, Vivek
通讯作者：
Malhotra, Vivek
影响因子：
4.8
作者：
Kumagai, Keigo;Kawano, Miyuki;Hanada, Kentaro
通讯作者：
Hanada, Kentaro
影响因子：
21.3
作者：
Litvak, V;Dahan, N;Lev, S
通讯作者：
Lev, S
影响因子：
4
作者：
Hausser, Angelika;Link, Gisela;Pfizenmaier, Klaus
通讯作者：
Pfizenmaier, Klaus
DOI：
10.1083/jcb.200612017
发表时间：
2007-07-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fugmann T;Hausser A;Schöffler P;Schmid S;Pfizenmaier K;Olayioye MA
通讯作者：
Olayioye MA