Reconstitution of β-adrenergic regulation of CaV1.2: Rad-dependent and Rad-independent protein kinase A mechanisms
Reconstitution of β-adrenergic regulation of CaV1.2: Rad-dependent and Rad-independent protein kinase A mechanisms
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DOI：
10.1073/pnas.2100021118
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发表时间：
2021-05-25
影响因子：
11.1
通讯作者：
Dascal, Nathan
中科院分区：
文献类型：
--
作者：
Katz, Moshe;Subramaniam, Suraj;Dascal, Nathan
L 型电压门控 CaV1.2 通道至关重要地调节心肌收缩。 0-肾上腺素能受体 (0-AR) 的激活通过蛋白激酶 A (PKA) 诱导的 CaV1.2 通道钙流入增加来增强收缩。迄今为止，完整的 0-AR 级联从未被异源重组。最近的一项研究发现 Rad（一种 CaV1.2 抑制蛋白）对于 PKA 调节 CaV1.2 至关重要。我们证实了这一发现，并重建了非洲爪蟾卵母细胞中 01-AR 或 02-AR 激动剂激活的完整途径。我们发现并区分了 CaV1.2 的 PKA 调节的两种不同途径：Rad 依赖性（类似于总数的 80%）和 Rad 依赖性。重建的系统再现了心肌细胞中 0-AR 调节的已知特征，并揭示了几个方面：翻译后修饰的 CaV1.2 变体的差异调节以及 01-AR 与 02-AR 活性的独特特征。该系统可以解决 0-AR-CaV1.2 级联中未解决的核心问题，并将促进儿茶酚胺诱导的心脏病疗法的开发。
L-type voltage-gated CaV1.2 channels crucially regulate cardiac muscle contraction. Activation of 0-adrenergic receptors (0-AR) augments contraction via protein kinase A (PKA)-induced increase of calcium influx through CaV1.2 channels. To date, the full 0-AR cascade has never been heterologously reconstituted. A recent study identified Rad, a CaV1.2 inhibitory protein, as essential for PKA regulation of CaV1.2. We corroborated this finding and reconstituted the complete pathway with agonist activation of 01-AR or 02-AR in Xenopus oocytes. We found, and distinguished between, two distinct pathways of PKA modulation of CaV1.2: Rad dependent (similar to 80% of total) and Rad independent. The reconstituted system reproduces the known features of 0-AR regulation in cardiomyocytes and reveals several aspects: the differential regulation of posttranslationally modified CaV1.2 variants and the distinct features of 01-AR versus 02-AR activity. This system allows for the addressing of central unresolved issues in the 0-AR-CaV1.2 cascade and will facilitate the development of therapies for catecholamine-induced cardiac pathologies.