Common components of patch-clamp internal recording solutions can significantly affect protein kinase A activity.
Common components of patch-clamp internal recording solutions can significantly affect protein kinase A activity.
复制标题
膜片钳内部记录溶液的常见成分可显着影响蛋白激酶 A 活性。
DOI：
10.1016/s0006-8993(99)01306-2
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发表时间：
1999
期刊：
影响因子：
2.9
通讯作者：
Lucero,MT
中科院分区：
文献类型：
--
作者：
Vargas,G;Yeh,TY;Blumenthal,DK;Lucero,MT
在体外和膜片钳实验中测试了全细胞内部记录溶液的常见成分对 cAMP 依赖性蛋白激酶活性的影响。氟化钾 (KF)、440 mM 三甲胺氯化物和排除牛血清白蛋白 (BSA) 降低了酶的活性，而乙二醇-双 (β-氨基乙基醚) N,N,N',N'-四乙酸 (EGTA) 和天冬氨酸钾盐、葡萄糖酸钾、硫酸甲酯和磷酸二氢钾盐则增加了酶的活性。向内部溶液中添加 KF 会导致 Ih 通道激活的 V1/2 发生超极化转变，这与 KF 诱导的蛋白激酶 A 活性降低一致。因此，在通过膜片钳技术研究通道生理学时，需要考虑内部溶液的组成。
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 (β-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 V1/2of Ihchannel 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.