膜片钳内液记录溶液的常见成分可显著影响蛋白激酶A的活性。
Common components of patch-clamp internal recording solutions can significantly affect protein kinase A activity.
作者信息
Vargas G, Yeh T Y, Blumenthal D K, Lucero M T
机构信息
Department of Physiology, University of Utah, School of Medicine, Salt Lake City, UT 84108, USA.
出版信息
Brain Res. 1999 May 15;828(1-2):169-73. doi: 10.1016/s0006-8993(99)01306-2.
Abstract
Common components of whole-cell internal recording solutions were tested both in vitro and in patch-clamp experiments for their effects on the activity of cAMP-dependent protein kinase. Potassium fluoride (KF), 440 mM trimethylamine chloride and exclusion of bovine serum albumin (BSA) decreased the activity of the enzyme, while ethylene glycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) and the potassium salts of aspartate, gluconate, methylsulfate and monobasic phosphate increased its activity. Addition of KF to the internal solution produced a hyperpolarizing shift in the V1/2 of Ih channel activation, consistent with the KF-induced reduction of protein kinase A activity. Therefore, consideration of the composition of internal solutions is warranted when studying channel physiology by patch-clamp techniques.
摘要
在体外和膜片钳实验中测试了全细胞内记录溶液的常见成分对环磷酸腺苷依赖性蛋白激酶活性的影响。440 mM 氟化钾（KF）、三甲氯化铵和排除牛血清白蛋白（BSA）会降低该酶的活性，而乙二醇双（β-氨基乙醚）N,N,N',N'-四乙酸（EGTA）以及天冬氨酸钾、葡萄糖酸钾、甲磺酸钾和磷酸二氢钾会增加其活性。向内部溶液中添加 KF 会使 Ih 通道激活的 V1/2 发生超极化偏移，这与 KF 诱导的蛋白激酶 A 活性降低一致。因此，在用膜片钳技术研究通道生理学时，有必要考虑内部溶液的成分。
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Anal Biochem. 1999 Feb 1;267(1):235-8. doi: 10.1006/abio.1998.2985.
2
Dopamine modulates inwardly rectifying hyperpolarization-activated current (Ih) in cultured rat olfactory receptor neurons.多巴胺调节培养的大鼠嗅觉受体神经元中的内向整流超极化激活电流（Ih）。
J Neurophysiol. 1999 Jan;81(1):149-58. doi: 10.1152/jn.1999.81.1.149.
3
Identification of a gene encoding a hyperpolarization-activated pacemaker channel of brain.一种编码大脑超极化激活起搏器通道的基因的鉴定。
Cell. 1998 May 29;93(5):717-29. doi: 10.1016/s0092-8674(00)81434-8.
4
Cross-modulation of glycine-activated Cl- channels by protein kinase C and cAMP-dependent protein kinase in the rat.蛋白激酶C和环磷酸腺苷依赖性蛋白激酶对大鼠甘氨酸激活的氯离子通道的交叉调节作用
J Physiol. 1998 Jan 15;506 ( Pt 2)(Pt 2):331-9. doi: 10.1111/j.1469-7793.1998.331bw.x.
5
Dopaminergic modulation of sodium current in hippocampal neurons via cAMP-dependent phosphorylation of specific sites in the sodium channel alpha subunit.多巴胺能通过对钠通道α亚基特定位点的环磷酸腺苷（cAMP）依赖性磷酸化作用来调节海马神经元中的钠电流。
J Neurosci. 1997 Oct 1;17(19):7330-8. doi: 10.1523/JNEUROSCI.17-19-07330.1997.
6
cAMP-dependent regulation of cardiac L-type Ca2+ channels requires membrane targeting of PKA and phosphorylation of channel subunits.环磷酸腺苷（cAMP）依赖性对心脏L型钙通道的调节需要蛋白激酶A（PKA）定位于细胞膜以及通道亚基的磷酸化。
Neuron. 1997 Jul;19(1):185-96. doi: 10.1016/s0896-6273(00)80358-x.
7
PKA-dependent regulation of mKv1.1, a mouse Shaker-like potassium channel gene, when stably expressed in CHO cells.当在CHO细胞中稳定表达时，蛋白激酶A对小鼠类Shaker钾通道基因mKv1.1的依赖性调节。
J Neurosci. 1993 Dec;13(12):5242-50. doi: 10.1523/JNEUROSCI.13-12-05242.1993.
8
Anchoring of protein kinase A is required for modulation of AMPA/kainate receptors on hippocampal neurons.蛋白激酶A的锚定是调节海马神经元上的AMPA/海人酸受体所必需的。
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9
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J Physiol. 1989 Aug;415:233-49. doi: 10.1113/jphysiol.1989.sp017720.
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