Properties of acetylcholinesterase in axolemma-enriched fractions isolated from bovine splenic nerve.
The properties of acetylcholinesterase (AChE) in axolemma-enriched fractions (AEF) from bovine splenic nerve were investigated to see if they differed in any way from those of the AChE in diaphragm muscle. The axolemmal enzyme had a low Km for acetylthiocholine (ca. 90 microM), exhibited substrate inhibition, and had a well-defined optimum of substrate concentration of 1 mM. The rate of hydrolysis of substrate decreased with increasing acyl chain length (acetyl- greater than propionyl- greater than butyryl-). The AChE inhibitors eserine and hexamethonium were competitive inhibitors of the membrane-bound enzyme, whereas lidocaine was a noncompetitive inhibitor; these results were comparable to the effect of these inhibitors on diaphragm muscle AChE. The axolemmal enzyme was more efficiently solubilized and more stable in nonionic detergents such as Triton X-100 and Tween 20 than charged detergents such as lysolecithin and zwitterionic detergents. These results indicate that the AChE present in bovine splenic nerve AEF is identical to the previously characterized AChE from other sources.
Hannesson
H H
HH
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University 23298-0614.
DeVries
G H
GH
D000110:Acetylcholinesterase / Q000302:isolation & purification* / Q000378:metabolism; D000122:Acetylthiocholine / Q000378:metabolism; D000818:Animals; D001369:Axons / Q000201:enzymology; D002417:Cattle; D003902:Detergents; D003964:Diaphragm / Q000201:enzymology; D018738:Hexamethonium; D006584:Hexamethonium Compounds / Q000494:pharmacology; D007700:Kinetics; D008012:Lidocaine / Q000494:pharmacology; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology; D010830:Physostigmine / Q000494:pharmacology; D012995:Solubility; D013154:Spleen / Q000294:innervation*; D013379:Substrate Specificity
D003902:Detergents; D006584:Hexamethonium Compounds; D008565:Membrane Proteins; D018738:Hexamethonium; D000122:Acetylthiocholine; D008012:Lidocaine; D010830:Physostigmine; D000110:Acetylcholinesterase