In vivo regulation of glycogen synthase kinase 3β activity in neurons and brains.
In vivo regulation of glycogen synthase kinase 3β activity in neurons and brains.
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DOI：
10.1038/s41598-017-09239-5
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发表时间：
2017-08-17
影响因子：
4.6
通讯作者：
Hisanaga SI
中科院分区：
文献类型：
--
作者：
Krishnankutty A;Kimura T;Saito T;Aoyagi K;Asada A;Takahashi SI;Ando K;Ohara-Imaizumi M;Ishiguro K;Hisanaga SI
糖原合成酶激酶3β (GSK3β)是一种多功能蛋白激酶，参与许多细胞活动，包括发育、分化和疾病。GSK3β被认为在Tyr216位点被自磷酸化激活，在Ser9位点被磷酸化失活。GSK3β的活性之前已经通过抑制Ser9磷酸化来评估，但它并不一定表明激酶本身的活性。在这里，我们应用Phos-tag SDS-PAGE技术分析了细胞和大脑中的GSK3β磷同型。GSK3β有3种磷酸同型；Ser9和Tyr216的双磷酸化，Tyr216的单磷酸化和非磷酸化同型。在培养细胞中Tyr216位点磷酸化的活性GSK3β占GSK3β总量的一半或更多。虽然胰岛素治疗增加了磷酸化Ser9的水平，但Ser9磷酸化仅发生在GSK3β的一小部分。在小鼠大脑中，GSK3β主要以活性形式存在，Ser9磷酸化很少，并且GSK3β的磷酸化同型随大脑区域、年龄、性别和疾病状况而变化。这些结果表明，Phos-tag SDS-PAGE方法提供了一种简单、合适的体内GSK3β活性测定方法，其活性受Ser9磷酸化以外的机制调控。
Glycogen synthase kinase 3β (GSK3β) is a multifunctional protein kinase involved in many cellular activities including development, differentiation and diseases. GSK3β is thought to be constitutively activated by autophosphorylation at Tyr216 and inactivated by phosphorylation at Ser9. The GSK3β activity has previously been evaluated by inhibitory Ser9 phosphorylation, but it does not necessarily indicate the kinase activity itself. Here, we applied the Phos-tag SDS-PAGE technique to the analysis of GSK3β phosphoisotypes in cells and brains. There were three phosphoisotypes of GSK3β; double phosphorylation at Ser9 and Tyr216, single phosphorylation at Tyr216 and the nonphosphorylated isotype. Active GSK3β with phosphorylation at Tyr216 represented half or more of the total GSK3β in cultured cells. Although levels of phospho-Ser9 were increased by insulin treatment, Ser9 phosphorylation occurred only in a minor fraction of GSK3β. In mouse brains, GSK3β was principally in the active form with little Ser9 phosphorylation, and the phosphoisotypes of GSK3β changed depending on the regions of the brain, age, sex and disease conditions. These results indicate that the Phos-tag SDS-PAGE method provides a simple and appropriate measurement of active GSK3β in vivo, and the activity is regulated by the mechanism other than phosphorylation on Ser9.
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影响因子：
34.7
作者：
Hur, Eun-Mi;Zhou, Feng-Quan
通讯作者：
Zhou, Feng-Quan
影响因子：
4.8
作者：
Jensen, Jorgen;Brennesvik, Erlend O.;Shepherd, Peter R.
通讯作者：
Shepherd, Peter R.
影响因子：
7
作者：
Hosokawa, Tomohisa;Saito, Taro;Hisanaga, Shin-ichi
通讯作者：
Hisanaga, Shin-ichi
影响因子：
4.8
作者：
Clodfelder-Miller, B;De Sarno, P;Jope, RS
通讯作者：
Jope, RS
DOI：
10.1073/pnas.93.7.2719
发表时间：
1996-04-02
影响因子：
11.1
作者：
Hoshi, M;Takashima, A;Imahori, K
通讯作者：
Imahori, K