Properties of acetylcholinesterase reconstituted in liposomes of a different charge.
Acetylcholinesterase (AChE) purified from mouse brain was reconstituted in liposomes of a different charge, and the properties of liposome-associated AChE were investigated. Relative to the Km value (38.5 microM) of AChE bound to a neutral liposome, the value of AChE reconstituted in a negatively-charged liposome decreased to 23.3 microM, whereas that of AChE in a positively-charged liposome increased to 90.9 microM. Additionally, AChE bound to a positively-charged liposome expressed a wider range of optimum pH than the enzyme in a negatively-charged liposome. In a stability study, it was found that soluble AChE was unstable at pH 5.5 and 7.4, while it was relatively stable at pH 10. Noteworthy, the immobilization of AChE to liposome enhanced the stability of soluble enzyme at acidic and neutral pH. Moreover, in the stabilization of the enzyme, a neutral liposome was more effective than charged liposomes, of which a positively-charged liposome was more effective than a negatively-charged liposome at acidic pH. Based on these results, it is proposed that while the Km value and the pH dependence of AChE activity are affected by the charge of liposome, the stability of AChE is determined mainly by a hydrophobic binding to a phospholipid membrane.
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Advanced Technology Research Center (#6-7), Agency for Defense Development, Yuseong, Taejon, Korea.
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D015060:1,2-Dipalmitoylphosphatidylcholine; D000110:Acetylcholinesterase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004134:Dimyristoylphosphatidylcholine; D005260:Female; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D013816:Thermodynamics
D008081:Liposomes; D015060:1,2-Dipalmitoylphosphatidylcholine; D000110:Acetylcholinesterase; D004134:Dimyristoylphosphatidylcholine