Reconstitution of beta-adrenergic modulation of large conductance, calcium-activated potassium (maxi-K) channels in Xenopus oocytes. Identification of the camp-dependent protein kinase phosphorylation site.
Reconstitution of beta-adrenergic modulation of large conductance, calcium-activated potassium (maxi-K) channels in Xenopus oocytes. Identification of the camp-dependent protein kinase phosphorylation site.
复制标题
非洲爪蟾卵母细胞中大电导钙激活钾 (maxi-K) 通道的 β-肾上腺素能调节的重建。
DOI：
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发表时间：
1998
影响因子：
4.8
通讯作者：
M. Kotlikoff
中科院分区：
文献类型：
--
作者：
Masayuki Nara;P. Dhulipala;Y. Wang;M. Kotlikoff
将人类大电导钙激活钾 (maxi-K) 通道（α 和 β 亚基）和 β2 肾上腺素受体基因在非洲爪蟾卵母细胞中共表达，以研究 β 肾上腺素能调节通道功能的机制。异丙肾上腺素和毛喉素使表达受体和两个通道亚基的电压钳位卵母细胞中的 maxi-K 钾通道电流分别增加 33 +/- 5% 和 35 +/- 8%，而不影响电流激活或失活。共表达α和β亚基的卵母细胞与仅表达α亚基的卵母细胞中异丙肾上腺素和毛喉素的刺激百分比没有差异，这表明α亚基是调节的目标。异丙肾上腺素的刺激作用几乎被细胞内注射环 AMP 依赖性蛋白激酶 (cAMP-PK) 调节亚基完全阻断，而注射环 GMP 依赖性蛋白激酶抑制肽几乎没有作用，表明 β2-肾上腺素能受体与 maxi-K 通道的细胞偶联涉及内源性 cAMP-PK。 α 亚基上几个潜在共有 cAMP-PK 磷酸化位点之一（丝氨酸 869）的突变几乎完全抑制 β-肾上腺素能受体/通道刺激耦合，而毛喉素仍然适度刺激电流（16 +/- 4%）。这些数据表明，β2 受体和 maxi-K 通道之间的生理耦合是通过 cAMP-PK 介导的通道 α 亚基上丝氨酸 869 的磷酸化而发生的。
The human large conductance, calcium-activated potassium (maxi-K) channel (alpha and beta subunits) and beta2-adrenergic receptor genes were coexpressed in Xenopus oocytes in order to study the mechanism of beta-adrenergic modulation of channel function. Isoproterenol and forskolin increased maxi-K potassium channel currents in voltage-clamped oocytes expressing the receptor and both channel subunits by 33 +/- 5% and 35 +/- 8%, respectively, without affecting current activation or inactivation. The percentage of stimulation by isoproterenol and forskolin was not different in oocytes coexpressing the alpha and beta subunits versus those expressing the only the alpha subunit, suggesting that the alpha subunit is the target for regulation. The stimulatory effect of isoproterenol was almost completely blocked by intracellular injection of the cyclic AMP dependent protein kinase (cAMP-PK) regulatory subunit, whereas injection of a cyclic GMP dependent protein kinase inhibitory peptide had little effect, indicating that cellular coupling of beta2-adrenergic receptors to maxi-K channels involves endogenous cAMP-PK. Mutation of one of several potential consensus cAMP-PK phosphorylation sites (serine 869) on the alpha subunit almost completely inhibited beta-adrenergic receptor/channel stimulatory coupling, whereas forskolin still stimulated currents moderately (16 +/- 4%). These data demonstrate that physiological coupling between beta2 receptors and maxi-K channels occurs by the cAMP-PK mediated phosphorylation of serine 869 on the alpha subunit on the channel.
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DOI：
10.1152/ajpcell.1993.265.1.c299
发表时间：
1993-07
期刊：
The American journal of physiology
影响因子：
--
作者：
B. E. Robertson;R. Schubert;Jorgen Hescheler;M. Nelson
通讯作者：
B. E. Robertson;R. Schubert;Jorgen Hescheler;M. Nelson
影响因子：
3.5
作者：
PALLANCK, L;GANETZKY, B
通讯作者：
GANETZKY, B
蛋白激酶 A 和磷酸酶抑制剂对 Ca(2) 激活的 K 通道的调节。
DOI：
10.1152/ajpcell.1991.261.2.c387
发表时间：
1991
期刊：
The American journal of physiology
影响因子：
--
作者：
Carl,A;Kenyon,JL;Uemura,D;Fusetani,N;Sanders,KM
通讯作者：
Sanders,KM
DOI：
10.1073/pnas.93.17.9200
发表时间：
1996-08-20
影响因子：
11.1
作者：
TsengCrank, J;Godinot, N;Reinhart, PH
通讯作者：
Reinhart, PH
影响因子：
--
作者：
A. L. Goldin
通讯作者：
A. L. Goldin