The Amino-terminal Portion of the JAK2 Protein Kinase Is Necessary For Binding and Phosphorylation of the Granulocyte-Macrophage Colony-stimulating Factor Receptor β Chain (*)
The Amino-terminal Portion of the JAK2 Protein Kinase Is Necessary For Binding and Phosphorylation of the Granulocyte-Macrophage Colony-stimulating Factor Receptor β Chain (*)
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JAK2 蛋白激酶的氨基末端部分对于粒细胞-巨噬细胞集落刺激因子受体 β 链的结合和磷酸化是必需的 (*)
DOI：
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发表时间：
1995
影响因子：
4.8
通讯作者：
A. Kraft
中科院分区：
文献类型：
--
作者：
Yanming Zhao;F. Wagner;S. Frank;A. Kraft
粒细胞-巨噬细胞集落刺激因子（GM-CSF）与其受体结合可刺激JAK2蛋白激酶活化、蛋白磷酸化以及JAK2与GM-CSF受体βc链的结合。为了更好地理解JAK2的不同结构域如何调节βc受体的结合和磷酸化，已经确定了JAK2结合所需的βc受体的最小部分。利用谷胱甘肽s -转移酶（GST）融合蛋白表达βc链膜近端结构域的不同部分，我们发现JAK2与458-495氨基酸结合，但与含有483- 559,483 -530或458-484氨基酸的融合蛋白结合很少。GST-βc 458-495与野生型（WT） JAK2结合同样良好，JAK2的羧基末端缺失去除蛋白激酶结构域（氨基酸1000-1129），激酶样结构域（氨基酸523-746）缺失。然而，氨基末端JAK2的缺失（氨基酸2-239）显著降低了与GST-βc的结合。Far Western blotting证实含有JAK2氨基酸1-294的GST融合蛋白与GST-βc 458-559结合，而不含有含有氨基酸295-522、523-746或747-1127的融合蛋白。将JAK2的WT和缺失与GM-CSF受体的α和βc亚基一起短暂表达，并对细胞进行GM-CSF处理，得到以下结果：1)WT JAK2以GM-CSF依赖的方式磷酸化βc亚基，2)激酶样结构域缺失磷酸化βc亚基，3)激酶结构域缺失和氨基末端缺失都不能刺激βc亚基的磷酸化。因此，βc亚基的磷酸化需要通过其氨基末端与JAK2结合。
The binding of granulocyte-macrophage colony-stimulating factor (GM-CSF) to its receptor stimulates JAK2 protein kinase activation, protein phosphorylation, and JAK2 association with the βc chain of the GM-CSF receptor. To better understand how different domains of the JAK2 function to regulate association and phosphorylation of the βc receptor, the minimal portion of the βc receptor necessary for JAK2 binding has been determined. Using glutathione S-transferase (GST) fusion proteins expressing different portions of the membrane-proximal domain of the βc chain, we demonstrate that JAK2 binds to amino acids 458-495, but showed little binding to fusion proteins containing amino acids 483-559, 483-530, or 458-484. The GST-βc 458-495 bound equally well to the wild type (WT) JAK2, a carboxyl-terminal deletion of JAK2 removing the protein kinase domain (amino acids 1000-1129), and a deletion of the kinase-like domain (amino acids 523-746). However, an amino-terminal JAK2 deletion (amino acids 2-239) markedly reduced binding to this GST-βc. Far Western blotting demonstrated that a GST fusion protein containing amino acids 1-294 of JAK2, but not fusion proteins containing amino acids 295-522, 523-746, or 747-1127, bound GST-βc 458-559. When the JAK2 WT and deletions were transiently expressed along with the α and βc subunits of the GM-CSF receptor and the cells were treated with GM-CSF, the following results were obtained: 1) WT JAK2 phosphorylated the βc subunit in a GM-CSF- dependent manner, 2) the kinase-like domain deletion phosphorylated the βc subunit, and 3) both the kinase domain deletion and the amino-terminal deletion failed to stimulate phosphorylation of the βc subunit. Therefore, phosphorylation of the βc subunit requires the binding of JAK2 through its amino terminus.
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DOI：
10.1073/pnas.91.23.11075
发表时间：
1994-11-08
影响因子：
11.1
作者：
CHINKERS, M
通讯作者：
CHINKERS, M
影响因子：
8
作者：
I. Firmbach-Kraft;M. Byers;Shows Tb;R. Dalla-Favera;John J. Krolewski
通讯作者：
I. Firmbach-Kraft;M. Byers;Shows Tb;R. Dalla-Favera;John J. Krolewski
激酶缺陷形式的 Jak2 抑制促红细胞生成素诱导的有丝分裂。
DOI：
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发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Zhuang,H;Patel,SV;He,TC;Sonsteby,SK;Niu,Z;Wojchowski,DM
通讯作者：
Wojchowski,DM
绘制细胞生长调节所需的α粒细胞-巨噬细胞集落刺激因子受体的胞质内区域。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Polotskaya,A;Zhao,Y;Lilly,MB;Kraft,AS
通讯作者：
Kraft,AS
DOI：
10.1073/pnas.88.2.627
发表时间：
1991-01-01
影响因子：
11.1
作者：
MAYER, BJ;JACKSON, PK;BALTIMORE, D
通讯作者：
BALTIMORE, D