Use of site-directed antibodies to probe the topography of the alpha 2 subunit of voltage-gated Ca2+ channels.
Polyclonal antibodies were raised against peptides corresponding to residues 1-15, 469-483 and 933-951 of the rabbit skeletal muscle L-type calcium channel alpha 2/delta primary translation product, for use as topological probes. Immunocytochemical comparison of the abilities of the antibodies to bind to the alpha 2 and delta subunits in intact and detergent-permeabilised rat dorsal root ganglion cells enabled the membrane orientation of these regions to be established. The resultant data indicate that the regions containing residues 1-15 and 469-483 of the alpha 2 subunit, and residues 1-17 of the delta subunit, are exposed on the extracellular surface of the membrane, findings consistent with a model that proposes alpha 2 to be entirely extracellular.
Brickley
K
K
Department of Pharmacology, Royal Free Hospital School of Medicine, (University of London), UK.
Campbell
V
V
Berrow
N
N
Leach
R
R
Norman
R I
RI
Wray
D
D
Dolphin
A C
AC
Baldwin
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D015220:Calcium Channels / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D005727:Ganglia, Spinal / Q000378:metabolism; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D015220:Calcium Channels; D010455:Peptides