Amphibian ryanodine receptor isoforms are related to those of mammalian skeletal or cardiac muscle.
The ryanodine receptor (RyR)-Ca2+ release channels of frog skeletal muscle have been purified as 30S protein complexes comprised of two high molecular weight polypeptides. The upper and lower bands of the frog doublet comigrated on sodium dodecyl sulfate polyacylamide gels with the mammalian skeletal and cardiac RyR polypeptides, respectively. Immunoblot analysis showed that a polyclonal antiserum to the rat skeletal RyR preferentially cross-reacted with the upper band, whereas monoclonal antibodies to the canine cardiac RyR preferentially cross-reacted with the lower band of the frog receptor doublet. Immunoprecipitation studies indicated the presence of two homooligomer 30S RyR complexes comprised of either the lower or upper polypeptide band of the frog doublet, and immunocytochemical staining revealed their colocalization in frog gastrocnemius muscle. After planar lipid bilayer reconstitution of the 30S frog RyR, single-channel currents were observed that exhibited a Na+ and Ca2+ conductance and pharmacological characteristics similar to those of the mammalian skeletal and cardiac Ca2+ release channels. These results suggest that amphibian skeletal muscle expresses two distinct RyR isoforms that share epitopes in common with the mammalian skeletal or cardiac RyR.
Lai
F A
FA
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill 27599-7260.
Liu
Q Y
QY
Xu
L
L
el-Hashem
A
A
Kramarcy
N R
NR
Sealock
R
R
Meissner
G
G
D000818:Animals; D015220:Calcium Channels / Q000378:metabolism; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D007536:Isomerism; D009132:Muscles / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D011233:Precipitin Tests; D011894:Rana pipiens / Q000378:metabolism*; D051381:Rats; D011950:Receptors, Cholinergic / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
D015220:Calcium Channels; D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel