Molecular interactions of the junctional foot protein and dihydropyridine receptor in skeletal muscle triads.
Isolated triadic proteins were employed to investigate the molecular architecture of the triad junction in skeletal muscle. Immunoaffinity-purified junctional foot protein (JFP), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), aldolase and partially purified dihydropyridine (DHP) receptor were employed to probe protein-protein interactions using affinity chromatography, protein overlay and crosslinking techniques. The JFP, an integral protein of the sarcoplasmic reticulum (SR) preferentially binds to GAPDH and aldolase, peripheral proteins of the transverse (T)-tubule. No direct binding of JFP to the DHP receptor was detected. The interactions of JFP with GAPDH and aldolase appear to be specific since other glycolytic enzymes associated with membranes do not bind to the JFP. The DHP receptor, an integral protein of the T-tubule, also binds GAPDH and aldolase. A ternary complex between the JFP and the DHP receptor can be formed in the presence of GAPDH. In addition, the DHP receptor binds to a previously undetected Mr 95 K protein which is distinct from the SR Ca2+ pump and phosphorylase b. The Mr 95 K protein is an integral protein of the junctional domain of the SR terminal cisternae. It is also present in the newly identified "strong triads" (accompanying paper). From these findings, we propose a new model for the triad junction.
Brandt
N R
NR
Department of Pharmacology, University of Miami School of Medicine, Florida 33101.
Caswell
A H
AH
Wen
S R
SR
Talvenheimo
J A
JA
D000818:Animals; D002121:Calcium Channel Blockers / Q000378:metabolism; D015220:Calcium Channels; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D066298:In Vitro Techniques; D008954:Models, Biological; D009124:Muscle Proteins / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism* / Q000648:ultrastructure; D011817:Rabbits; D011950:Receptors, Cholinergic / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D009124:Muscle Proteins; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; D019837:Ryanodine Receptor Calcium Release Channel; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005634:Fructose-Bisphosphate Aldolase