Identification and localization of two triad junctional foot protein isoforms in mature avian fast twitch skeletal muscle.
We report evidence for two foot protein isoforms in chicken pectoral muscle. (i) Two polypeptides with molecular masses of approximately 500 kDa copurify with [3H]ryanodine binding. (ii) Both polypeptides are associated with oligomeric proteins similar in size to the mammalian skeletal muscle foot protein. (iii) The polypeptides are shown to be unique by limited proteolysis. (iv) By using isoform-specific antibodies, the polypeptides are shown to be subunits of different [3H]ryanodine-binding proteins. Using immunolabeling techniques, we have localized these proteins in chicken breast muscle by both light and electron microscopy. (v) From immunofluorescent light microscopy of longitudinal sections, it was determined that both ryanodine-binding protein isoforms exhibit identical repetitive punctate distributions near the Z-lines. (vi) In serial cross-sections both proteins have similar distributions in the same fibers. (vii) Both proteins were found to be associated with the terminal cisternae of the sarcoplasmic reticulum by immunoelectron microscopy. Based on their localization to the triadic junction, their large size and their ability to bind [3H]ryanodine, these proteins are identified as foot proteins. In conclusion, two distinct homo-oligomeric foot proteins coexist in avian fast twitch skeletal muscle. We have termed these proteins, alpha and beta foot proteins.
Airey
J A
JA
Department of Pharmacology and the Cell and Molecular Biology Program, University of Nevada, Reno 89557.
Beck
C F
CF
Murakami
K
K
Tanksley
S J
SJ
Deerinck
T J
TJ
Ellisman
M H
MH
Sutko
J L
JL
D000470:Alkaloids / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002499:Centrifugation, Density Gradient; D002645:Chickens; D007425:Intracellular Membranes / Q000378:metabolism*; D046911:Macromolecular Substances; D008861:Microsomes / Q000378:metabolism*; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000302:isolation & purification / Q000378:metabolism*; D012433:Ryanodine / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel
D000470:Alkaloids; D046911:Macromolecular Substances; D009124:Muscle Proteins; D011950:Receptors, Cholinergic; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine