Phosphorylation of Chromogranin A and Catecholamine Secretion Stimulated by Elevation of Intracellular Ca2+ in Cultured Bovine Adrenal Medullary Cells*

Phosphorylation of Chromogranin A and Catecholamine Secretion Stimulated by Elevation of Intracellular Ca2+ in Cultured Bovine Adrenal Medullary Cells*
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培养的牛肾上腺髓质细胞胞内 Ca2+ 升高刺激嗜铬蛋白 A 的磷酸化和儿茶酚胺分泌*

DOI:
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发表时间:
1996
影响因子:
4.8
通讯作者:
F. Izumi
F. Izumi
中科院分区:
生物学2区
文献类型:
--
作者:
N. Yanagihara;Y. Oishi;Hideyuki Yamamoto;M. Tsutsui;Jun Kondoh;Tutomu Sugiura;E. Miyamoto;F. Izumi

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我们最近从牛肾上腺髓质细胞中分离出了Ca 2 +/钙调蛋白依赖性蛋白激酶II(CaM激酶II)的新的70 kDa内源性底物(Yanagihara,N.,Toyohira,Y.,山本,H.,Ohta,Y.,Tsutsui,M.,Miyamoto,E.,和Izumi,F.(1994)Mol. Pharmacol. 46,423-430)。在这里,我们报告的70 kDa的蛋白质的序列分析,并检查其磷酸化的各种蛋白激酶在体外和去极化的培养细胞。蛋白质测序和免疫印迹显示,70 kDa的蛋白质是嗜铬粒蛋白A(CgA)或密切相关的蛋白质。部分纯化的CgA被环AMP依赖性蛋白激酶和蛋白激酶C以及CaM激酶II磷酸化。胰蛋白酶磷酸肽映射模式的CGA不同,这些蛋白激酶。在32 P-标记的牛肾上腺髓质细胞,56 mM K+增加磷酸化的CgA和儿茶酚胺分泌在类似的时间和浓度依赖性的方式,这两个都被抑制20 mM MgSO 4,电压依赖性钙通道的抑制剂。这些研究结果表明,CGA作为底物的几个多功能的蛋白激酶和细胞内Ca 2+的升高刺激CGA的磷酸化与儿茶酚胺分泌在培养的肾上腺髓质细胞。
We have recently isolated a new endogenous substrate of 70 kDa for Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) from bovine adrenal medullary cells (Yanagihara, N., Toyohira, Y., Yamamoto, H., Ohta, Y., Tsutsui, M., Miyamoto, E., and Izumi, F. (1994) Mol. Pharmacol. 46, 423-430). Here we report the sequence analysis of the 70-kDa protein and examine its phosphorylation by various protein kinases in vitro and by depolarization of the cultured cells. Protein sequencing and immunoblotting revealed that the 70-kDa protein is chromogranin A (CgA) or a closely related protein. Partially purified CgA was phosphorylated by cyclic AMP-dependent protein kinase and protein kinase C as well as CaM kinase II. Tryptic phosphopeptide mapping patterns of CgA differed among these protein kinases. In 32P-labeled bovine adrenal medullary cells, 56 mM K+ increased the phosphorylation of CgA and catecholamine secretion in similar time- and concentration-dependent manners, both of which were inhibited by 20 mM MgSO4, an inhibitor of voltage-dependent Ca2+ channels. These findings suggest that CgA serves as a substrate for several multifunctional protein kinases and that the elevation of the intracellular Ca2+ stimulates the phosphorylation of CgA associated with catecholamine secretion in cultured adrenal medullary cells.
DOI: 10.1016/s0021-9258(18)77318-3
发表时间: 1990-08
期刊: The Journal of biological chemistry
影响因子: --
作者:
S. Yoo;J. Albanesi
通讯作者: S. Yoo;J. Albanesi
DOI: 10.1016/s0021-9258(18)33559-2
发表时间: 1982-11
期刊: The Journal of biological chemistry
影响因子: --
作者:
J. Haycock;J. Meligeni;W. Bennett;J. Waymire
通讯作者: J. Haycock;J. Meligeni;W. Bennett;J. Waymire
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DOI: --
发表时间: 1988
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Schulman,H
DOI: --
发表时间: 1988
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Holz,RW
DOI: --
发表时间: 1984
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Frigon,RP