Subcellular distribution and characterization of acetylcholinesterase activities from sheep platelets: relationships between temperature-dependence and environment.
Acetylcholinesterase is a key enzyme in cholinergic neurotransmission for hydrolyzing acetylcholine and has been shown to possess arylacylamidase activity in addition to esterase activity. The enzyme is found at various loci, where its functional significance remains to be clarified, and it exists in multiple molecular forms. Sheep platelets have been shown to exhibit acetylcholinesterase activity associated with plasma membrane (Bp), endoplasmic reticulum (Cp), mitochondria granules (Dp), and soluble (As) fractions. These activities show differences in some physicochemical and kinetic properties. The soluble acetylcholinesterase is the most thermostable, and the enzyme from the Cp fractions shows the lowest affinity for the acetylthiocholine substrate and the strongest inhibition by fluoride. In all cases a noncompetitive inhibition of the enzyme by this ion is found. When membrane-bound acetylcholinesterases were assayed at temperatures between 12 degrees C and 33 degrees C, the Arrhenius plots of all activities exhibited a break point at about 17 degrees C. This discontinuity was abolished by addition of detergent to the assay medium (0.02% Triton X-100, final concentration). Their Hill coefficients were calculated in the presence of fluoride, showing unitary values in all cases, which points to a noncooperative effect and nonallosteric behavior in the particulate enzyme. These results suggest that the sheep platelet acetylcholinesterase associated with membrane-bound systems is modulated by the physical state of its environment, despite the fact that the enzyme might be lipid- or nonlipid-dependent.
Sánchez-Yagüe
J
J
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Salamanca, Spain.
Cabezas
J A
JA
Llanillo
M
M
D000110:Acetylcholinesterase / Q000097:blood* / Q000493:pharmacokinetics / Q000502:physiology; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology* / Q000502:physiology / Q000648:ultrastructure; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D015388:Organelles / Q000201:enzymology / Q000648:ultrastructure; D012756:Sheep; D013696:Temperature
D000110:Acetylcholinesterase