Reconstitution of the skeletal muscle dihydropyridine receptor. Functional interaction among alpha 1, beta, gamma and alpha 2 delta subunits.
The L-type voltage-dependent Ca2+ channel purified from skeletal muscle by virtue of its dihydropyridine (DHP) binding activity, is composed of alpha 1, alpha 2 delta, beta and gamma subunits. The alpha 1 subunit has the ability to function alone as a Ca2+ channel and a receptor for DHP and other Ca2+ channel antagonists. In this study, the non-alpha 1 components coexpressed with alpha 1 in COS cells were investigated for their effects on DHP binding and suppression of an anomalous allosteric regulation of the phenylalkylamine (-)D600 in complexes lacking one or more subunits. (-)D600 increased DHP binding to membranes of COS cells expressing alpha 1 beta while it did not affect DHP binding to skeletal muscle membranes. Coexpression of gamma or alpha 2 delta with alpha 1 beta partially suppressed this effect. Coexpression of all the subunits completely eliminated the stimulatory effect of (-)D600, while at the same time increasing the affinity of the complex for DHP to that stabilized in partial complexes by the phenylalkylamine. These results demonstrate that all of the components that co-purify are required for the formation of a functional DHP receptor having the properties of the native skeletal muscle DHP receptor.
Suh-Kim
H
H
Department of Anatomy, Ajou University, Suwon, Korea.
Wei
X
X
Klos
A
A
Pan
S
S
Ruth
P
P
Flockerzi
V
V
Hofmann
F
F
Perez-Reyes
E
E
Birnbaumer
L
L
D000818:Animals; D019556:COS Cells; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D020746:Calcium Channels, L-Type; D004095:Dihydropyridines / Q000378:metabolism; D015870:Gene Expression; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D004095:Dihydropyridines; D009124:Muscle Proteins; C038806:1,4-dihydropyridine