title: Involvement of calcium ions in the properties of cardiolipin-associated erythrocyte acetylcholinesterase.
abstract: Lipoprotein forms of acetylcholinesterase from bovine erythrocytes gave non-linear Arrhenius plots with a break at 20 degrees C and contained cardiolipin. The break in the Arrhenius plot was abolished by incubation of the enzyme in high salt (I = 1.8), but only in Ca2+ -chelating conditions. At I = 1.8 neither NaCl alone, CaCl2 nor sodium phosphate at acidic pH abolished the break. However, at this ionic strength either NaCl in 2 mM sodium phosphate (pH 7.4) or sodium phosphate, pH 8, or 1.0 M Na2CO3/NaHCO3 (pH 8.5--10, were able to remove the break. The Arrhenius plot break was regenerated by the addition of Ca2+ to the high salt-treated enzyme with mild homogenization, but could not be regenerated in the presence of EDTA unless CaCl2 was added in excess of the EDTA. Conditions which abolished the break enabled endogenous cardiolipin to be removed from the enzyme by chloroform/methanol extraction Cardiolipin from acetylcholinesterase incubated in high salt in Ca2+ -chelating conditions was not accessible to digestion by phospholipase A2, and was not separated from the enzyme by flotation in a sucrose density gradient or by Sephadex G-200 chromatography. Thus both Ca2+ and cardiolipin appear to be inaccessible, possibly by being tightly associated in the hydrophobic core of the enzyme by ionic and hydrophobic forces. Ca2+ may modulate the temperature dependence of acetylcholinesterase activity through a functionally linked ionic interaction with the enzyme-cardiolipin complex.
mesh_terms: D000110:Acetylcholinesterase / Q000097:blood*; D000818:Animals; D002118:Calcium / Q000097:blood*; D002308:Cardiolipins / Q000097:blood*; D002417:Cattle; D004912:Erythrocytes / Q000201:enzymology*; D009994:Osmolar Concentration; D011485:Protein Binding; D012997:Solvents; D013696:Temperature; D013816:Thermodynamics
chemical_list: D002308:Cardiolipins; D012997:Solvents; D000110:Acetylcholinesterase; D002118:Calcium