Reconstitution of β-adrenergic regulation of CaV1.2: Rad-dependent and Rad-independent protein kinase A mechanisms

Moshe Katz, Suraj Subramaniam, Orna Chomsky-Hecht, Vladimir Tsemakhovich, Veit Flockerzi, Enno Klussmann, Joel A. Hirsch, Sharon Weiss, Nathan Dascal

Research output: Contribution to journalArticlepeer-review

Abstract

L-type voltage-gated CaV1.2 channels crucially regulate cardiac muscle contraction. Activation of β-adrenergic receptors (β-AR) augments contraction via protein kinase A (PKA)–induced increase of calcium influx through CaV1.2 channels. To date, the full β-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 β1-AR or β2-AR in Xenopus oocytes. We found, and distinguished between, two distinct pathways of PKA modulation of CaV1.2: Rad dependent (∼80% of total) and Rad independent. The reconstituted system reproduces the known features of β-AR regulation in cardiomyocytes and reveals several aspects: the differential regulation of posttranslationally modified CaV1.2 variants and the distinct features of β1-AR versus β2-AR activity. This system allows for the addressing of central unresolved issues in the β-AR–CaV1.2 cascade and will facilitate the development of therapies for catecholamine-induced cardiac pathologies.

Original languageEnglish
Article numbere2100021118
JournalProceedings of the National Academy of Sciences of the United States of America
Volume118
Issue number21
DOIs
StatePublished - 25 May 2021

Keywords

  • Calcium channel
  • adrenergic
  • cardiac
  • heterologous
  • protein kinase A

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