Molecular forms of acetyl- and butyrylcholinesterase in normal and dystrophic mouse brain.
In searching for possible differences in the composition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) forms in dystrophic brain, the distribution of various enzyme molecules in normal (NB) and dystrophic (DB) 129B6F1/J mouse brain has been investigated. The tissue was sequentially extracted with saline (S1) and with saline-Triton X-100 buffers (S2) to release soluble and membrane-bound cholinesterases. About 15% of the AChE and 35% of the BuChE activities in NB were recovered in S1, and the rest in S2. G4, G2, and G1 AChE and BuChE forms were identified in the soluble fractions obtained from NB and DB. The shift in sedimentation values of the separated AChE and BuChE species in sucrose gradients made with and without detergents revealed the occurrence of hydrophilic (H) and amphiphilic (A) variants of cholinesterases in the extracts. The amphiphilic properties of the several AChE and BuChE molecules were analyzed by Triton X-114 phase-partitioning and by phenyl-agarose chromatography. A12 (1%), G4A (72%), G4H (8%), and G2A + G1A (19%) AChE molecules, and G4A (34%), G4H (19%), and G2A + G1A (47%) BuChE forms, were identified in NB. The G4A AChE and BuChE isoforms differed in their interaction with Triton X-114 and with a hydrophobic matrix. Neither the extent of cholinesterase solubilization, nor the distribution of individual enzyme forms, was significantly altered in DB. The lack of specific differences in the distribution of AChE and BuChE forms between NB and DB suggests that the biosynthetic pathway leading to the various enzyme forms is altered in muscle but not in dystrophic mouse brain.
Moral-Naranjo
M T
MT
Departamento de Bioquímica y Biología Molecular A, Universidad de Murcia, Spain.
Cabezas-Herrera
J
J
Vidal
C J
CJ
D000110:Acetylcholinesterase / Q000737:chemistry* / Q000302:isolation & purification; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002091:Butyrylcholinesterase / Q000737:chemistry* / Q000302:isolation & purification; D002499:Centrifugation, Density Gradient; D002847:Chromatography, Agarose; D007527:Isoenzymes / Q000737:chemistry* / Q000302:isolation & purification; D051379:Mice; D008815:Mice, Inbred Strains; D009137:Muscular Dystrophy, Animal / Q000201:enzymology*; D051381:Rats
D007527:Isoenzymes; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase