Amphiphilic and hydrophilic forms of acetyl- and butyrylcholinesterase in human brain.
Human brain acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were sequentially extracted, first with a Tris-saline buffer (S1) and then with 1% (w/v) Triton X-100 (S2). About 20 and 30% of the AChE and BuChE activities were recovered in S1 and most of the remaining enzymes in S2. Main molecular forms of about 10.5 S and 12.0 S, G4 forms of AChE and BuChE, and smaller amounts of 4.5 S and 5.5 S forms, G1 species of AChE and BuChE, were measured in S1. Application of Triton X-114 phase partitioning and affinity chromatography on phenyl-agarose to S1 revealed that 25% of the AChE and none of the BuChE molecules displayed amphiphilic properties. Analysis of the enzyme activity retained by the phenyl-agarose showed that G1 AChE constituted the bulk of the amphiphilic molecules released without detergent. Main G4 forms of AChE and BuChE were found in the S2 extract. Eighty and 45% of the AChE and BuChE activities in S2 were measured in the detergent-rich phase by Triton X-114 phase partitioning. Thus, most of the AChE and about half of the BuChE molecules in S2 displayed amphiphilic properties. The main peak of BuChE, a 12.0 S form in gradients made with Triton X-100, splits into two peaks of 9.5 S and 12.5 S in Brij 96-containing gradients. This suggests that hydrophilic G4 BuChE forms are predominant in S1 and that hydrophilic and amphiphilic isoforms coexist in S2.
Sáez-Valero
J
J
Departamento de Bioquímica y Biología Molecular-A, Universidad de Murcia, Spain.
Tornel
P L
PL
Muñoz-Delgado
E
E
Vidal
C J
CJ
D000110:Acetylcholinesterase / Q000737:chemistry*; D001921:Brain / Q000201:enzymology*; D002021:Buffers; D002091:Butyrylcholinesterase / Q000737:chemistry*; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D006801:Humans; D007202:Indicators and Reagents; D007527:Isoenzymes / Q000737:chemistry; D017830:Octoxynol; D013395:Sucrose
D002021:Buffers; D007202:Indicators and Reagents; D007527:Isoenzymes; D013395:Sucrose; D017830:Octoxynol; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase