Isolation from cholinergic synapses of a protein that binds to membranes in a calcium-dependent manner.
A protein that binds to membranes in a calcium-dependent manner between calcium concentrations of 10(-5) and 10(-6) M has been isolated in large amounts (20 mg/kg tissue) from the entirely cholinergic electric organ of Torpedo marmorata. The protein bound reversibly to membrane fractions in a calcium-specific and saturable manner. The protein also bound to lipids isolated from Torpedo electric organ and to clathrin-coated vesicles prepared from pig brain. The protein bound to a Triton X-100-sensitive site. It had an apparent subunit molecular weight of 32,000 by polyacrylamide gel electrophoresis and of 35,900 by amino acid analysis; a broad isoelectric range of 4.8 to 5.5; and 27% of its amino acids after hydrolysis were observed to be aspartic and glutamic acids. Synaptosomes derived from electric organ were enriched in the protein which is probably localised within the nerve ending. It was localised in the synaptic region of the electric organ by means of immunofluorescence. In the electric lobe, discrete patches of fluorescence were seen within the cell bodies that innervate the electric organ. The protein may be involved in the recognition of membranes within the cholinergic neurone. Proteins with similar purification properties were found in all tissues investigated so far, and polypeptides of subunit molecular weight 32,000 were identified in bovine adrenal medulla and guinea pig brain synaptosomes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017302:Annexins; D001921:Brain / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004557:Electric Organ / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D008970:Molecular Weight; D013552:Swine; D013569:Synapses / Q000187:drug effects / Q000378:metabolism*; D013574:Synaptosomes / Q000378:metabolism*; D014101:Torpedo
D000596:Amino Acids; D017302:Annexins; D002135:Calcium-Binding Proteins; D002118:Calcium