Cytosolic choline acetyltransferase binds specifically to cholinergic plasma membrane of rat brain synaptosomes to generate membrane-bound enzyme.
Uncovering the way membrane-bound choline acetyltransferase (ChAT) interacts with membranes and with which membrane in cholinergic neurons may help in understanding its role in acetylcholine metabolism. Subfractionation of rat hippocampal synaptosomes aiming to separate synaptic vesicles from plasma membranes shows that membrane-bound ChAT is bound to plasma membrane. Either detergents or urea and alkali can solubilize membrane-bound enzyme. Detergent-solubilized enzyme has a higher sedimentation rate than urea-alkali solubilized or cytosolic ChAT. Once dissociated, membrane-bound ChAT reassociates specifically with cholinergic plasma membranes, a process that was abolished by previous treatment of membranes with trypsin. Cytosolic ChAT behaves similarly. Thus, in cholinergic synaptosomes, ChAT exists as cytosolic and peripheral activity. Cytosolic ChAT generates peripheral enzyme most probably by interacting with a protein of plasma membrane of cholinergic nerve terminals. This "receptor" protein might regulate the amount of membrane-bound ChAT in cholinergic neurons.
Gabrielle
Pahud
P
Department of Pharmacology (APSIC), Centre Médical Universitaire, 1, rue Michel Servet, 1211 Geneva 4, Switzerland.
Jeana
Medilanski
M
Lorenza
Eder-Colli
EC
D000468:Alkalies / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002254:Carbonates / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology; D004338:Drug Combinations; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012995:Solubility; D012996:Solutions; D013347:Subcellular Fractions / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism*; D014018:Tissue Distribution; D014508:Urea / Q000494:pharmacology
D000468:Alkalies; D002254:Carbonates; D003902:Detergents; D004338:Drug Combinations; D012996:Solutions; D011092:Polyethylene Glycols; D014508:Urea; D017830:Octoxynol; C010615:Nonidet P-40; D002795:Choline O-Acetyltransferase