Induction of calcium currents by the expression of the alpha 1-subunit of the dihydropyridine receptor from skeletal muscle.
The dihydropyridine (DHP) receptor purified from skeletal muscle comprises five protein subunits (alpha 1, alpha 2, beta, gamma and delta) and produces Ca2+ currents that are blocked by DHPs. Cloning of the alpha 1- and alpha 2-subunits, the former affinity-labelled by DHP, has shown that the alpha 1-subunit is expressed in skeletal muscle alone, whereas the alpha 2- and delta- subunits are also expressed in other tissues. Although the transient expression of the alpha 1-subunit in myoblasts from dysgenic mice (but not in oocytes) has been demonstrated, the use of these expression systems to determine the function of the alpha 1- subunit is complicated by the presence of endogenous Ca2+ currents, which may reflect the constitutive expression of proteins similar to the alpha 2-, beta-, gamma- and/or delta-subunits. We therefore selected a cell line which has no Ca2+ currents or alpha 2- subunit, and probably no delta-subunit for stable transformation with complementary DNA of the alpha 1- subunit. The transformed cells express DHP-sensitive, voltage-gated Ca2+ channels, indicating that the minimum structure of these channels is at most an alpha 1 beta gamma complex and possibly an alpha 1- subunit alone.
Perez-Reyes
E
E
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
Kim
H S
HS
Lacerda
A E
AE
Horne
W
W
Wei
X Y
XY
Rampe
D
D
Campbell
K P
KP
Brown
A M
AM
Birnbaumer
L
L
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000345:Affinity Labels; D000818:Animals; D015220:Calcium Channels / Q000502:physiology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004095:Dihydropyridines / Q000494:pharmacology; D004553:Electric Conductivity; D005786:Gene Expression Regulation; D015151:Immunoblotting; D007739:L Cells; D046911:Macromolecular Substances; D051379:Mice; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis; D011978:Receptors, Nicotinic / Q000235:genetics / Q000502:physiology*; D014162:Transfection; D014170:Transformation, Genetic
D000345:Affinity Labels; D015220:Calcium Channels; D004095:Dihydropyridines; D046911:Macromolecular Substances; D011978:Receptors, Nicotinic; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; C038806:1,4-dihydropyridine; D004247:DNA