Structural heterogeneity and subcellular distribution of nicotinic synapse-associated proteins.
Peripheral membrane proteins (Mr = 43,000) are associated with Torpedo membranes highly enriched in nicotinic receptor. These 43,000-dalton proteins are not required for ion translocation or other known receptor functions, but they have been implicated in constraint of the nicotinic receptor within the plane of the membrane bilayer. Sodium dodecyl sulfate-polyacrylamide electrophoresis allows partial resolution of the 43,000-dalton band into a doublet. We have carried out further analysis using two-dimensional gel electrophoresis, which reveals the existence of at least seven Coomassie blue-staining spots in the isoelectric focusing dimension. Peptide maps of the individual spots serve to elucidate the observed electrophoretic complexity. Three different membrane-bound proteins, designated v1, v2, and v3, were identified on the basis of their characteristic peptide maps which show no apparent homology in amino acid sequence. Two of these proteins, v1 and v2, are resolved into multiple spots in the isoelectric focusing dimension, but each group of isoelectric focusing variants has nearly identical peptide fingerprints. Of relevance to the putative role of these proteins in synaptic or receptor supramolecular structures is the observation that only v1 is exclusively membrane bound, and co-purifies with receptor whereas both v2 and v3 are also prominent proteins of the cytoplasm and are depleted from membrane fractions most enriched in receptor. These proteins may interact in the formation or maintenance of synaptic and nicotinic receptor supramolecular structures.
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004557:Electric Organ / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000302:isolation & purification*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011950:Receptors, Cholinergic / Q000302:isolation & purification*; D011978:Receptors, Nicotinic / Q000302:isolation & purification*; D013569:Synapses / Q000378:metabolism*; D014101:Torpedo; D014357:Trypsin
D008565:Membrane Proteins; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; D014357:Trypsin