Ser-27, Tyr-10 and Tyr-7 in the alpha-chain of pig stomach H+,K(+)-ATPase as Ca(2+)-dependent phosphorylatable sites by intrinsic and extrinsic protein kinases.
Ser-27, Tyr-10 and Tyr-7 in the alpha-chain of pig stomach H+,K(+)-ATPase as Ca(2+)-dependent phosphorylatable sites by intrinsic and extrinsic protein kinases.
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猪胃 H ,K( )-ATP 酶 α 链中的 Ser-27、Tyr-10 和 Tyr-7 作为内部和外部蛋白激酶的 Ca(2 ) 依赖性磷酸化位点。
DOI：
10.1006/bbrc.1996.1589
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发表时间：
1996
影响因子：
3.1
通讯作者：
K. Taniguchi
中科院分区：
文献类型：
--
作者：
K. Togawa;S. Kaya;A. Shimada;T. Imagawa;S. Mårdh;J. Corbin;U. Kikkawa;K. Taniguchi
当猪胃膜H ~+，K ~（+）-ATP酶制剂与[γ-~（32）P]ATP，Mg ~（2+）和Ca ~（2+）孵育时，在H ~+，K ~（+）-ATP酶N-末端α链上的特异性Tyr和Ser残基发生可逆磷酸化，而在α链的其它区域没有任何可检测的磷酸化。温和的甲苯磺酰基苯丙氨酰氯甲基酮胰蛋白酶处理，然后通过反相柱色谱法得到三个放射性肽峰。第一个峰含有Tyr 10（32 P）和Tyr 7（32 P），第二个峰含有Tyr 10（32 P）。第三个峰含有Ser 27（32 P），它也是用蛋白激酶-C + Ca ~（2+）或蛋白激酶-A磷酸化的部分纯化的H ~+，K（+）-ATP酶制剂经胰蛋白酶处理后获得的。这是蛋白激酶对H+，K（+）-ATP酶α链的Ca ~（2+）依赖性磷酸化作用的首次证实。
When pig stomach membrane H+,K(+)-ATPase preparations were incubated with [gamma-32P]ATP, Mg2+ and Ca2+, reversible phosphorylation of specific Tyr and Ser residues in the N-terminal alpha-chain of H+,K(+)-ATPase occurred without any detectable phosphorylation in other regions of the alpha-chain. Mild tosylphenylalanyl chloromethyl ketone trypsin treatment followed by reverse-phase column chromatography yielded three radioactive peptide peaks. The first peak contained both Tyr10(32P) and Tyr7(32P) and the second peak contained Tyr10(32P). The third peak contained Ser27(32P) which was also obtained after trypsin treatment of partially purified H+,K(+)-ATPase preparations phosphorylated with protein kinase-C + Ca2+ or protein kinase-A. This is the first demonstration of Ca2(+)-dependent phosphorylation of the alpha-chain of H+,K(+)-ATPase by protein kinases.