Are soluble and membrane-bound rat brain acetylcholinesterase different?
Salt-soluble and detergent-soluble acetylcholinesterases (AChE) from adult rat brain were purified to homogeneity and studied with the aim to establish the differences existing between these two forms. It was found that the enzymatic activities of the purified salt-soluble AChE as well as the detergent-soluble AChE were dependent on the Triton X-100 concentration. Moreover, the interaction of salt-soluble AChE with liposomes suggests amphiphilic behaviour of this enzyme. Serum cholinesterase (ChE) did not bind to liposomes but its activity was also detergent-dependent. Detergent-soluble AChE remained in solution below critical micellar concentrations of Triton X-100. SDS polyacrylamide gel electrophoresis of purified, Biobeads-treated and iodinated detergent-soluble 11 S AChE showed, under non reducing conditions, bands of 69 kD, 130 kD and greater than 250 kD corresponding, respectively, to monomers, dimers and probably tetramers of the same polypeptide chain. Under reducing conditions, only a 69 kD band was detected. It is proposed that an amphiphilic environment stabilizes the salt-soluble forms of AChE in the brain in vivo and that detergent-soluble Biobeads-treated 11 S AChE possess hydrophobic domain(s) different from the 20 kD peptide already described.
Andres
C
C
Centre de Neurochimie du C.N.R.S., Strasbourg, France.
el Mourabit
M
M
Stutz
C
C
Mark
J
J
Waksman
A
A
D000110:Acetylcholinesterase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002802:Cholinesterases / Q000097:blood / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007457:Iodine Radioisotopes; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D008297:Male; D008566:Membranes / Q000201:enzymology; D008863:Microspheres; D017830:Octoxynol; D011092:Polyethylene Glycols; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D003902:Detergents; D007457:Iodine Radioisotopes; D007527:Isoenzymes; D008081:Liposomes; D011092:Polyethylene Glycols; D017830:Octoxynol; D000110:Acetylcholinesterase; D002802:Cholinesterases