Stable expression and coupling of cardiac L-type Ca2+ channels with beta 1-adrenoceptors.

Abstract:

:A number of neurotransmitters modulate cardiac dihydropyridine-sensitive L-type Ca2+ channels through several homologous G protein-coupled receptors. Previous studies that have examined receptor-Ca2+ channel interactions have suffered because of the coexpression of various receptor subtypes in native cells. To study the functional coupling of a particular receptor subtype to these channels, rabbit cardiac Ca2+ channel alpha 1 and skeletal beta and alpha 2/delta subunits were stably expressed in baby hamster kidney cells. In this stable cell line, Ca2+ channels remained at high levels (> 1000 fmol/mg protein, or 2700 channels per cell) over extended times. The expressed recombinant Ca2+ channels displayed the voltage dependence of activation and inactivation, unitary conductance, and pharmacology characteristic of native cardiac L-type Ca2+ channels. Subsequent coexpression of the beta 1-adrenoceptors (150 to 300 fmol/mg protein) with the Ca2+ channels resulted in cell responsiveness to the extracellular application of isoproterenol. These results indicate that heterogeneous expression in mammalian cells provides a useful system for studying both biophysical analysis of Ca2+ channel properties and receptor-coupled regulatory processes.

journal_name

Circ Res

journal_title

Circulation research

authors

Yatani A,Wakamori M,Niidome T,Yamamoto S,Tanaka I,Mori Y,Katayama K,Green S

doi

10.1161/01.res.76.3.335

subject

Has Abstract

pub_date

1995-03-01 00:00:00

pages

335-42

issue

3

eissn

0009-7330

issn

1524-4571

journal_volume

76

pub_type

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