Characterization of a tetrameric G4 form of acetylcholinesterase from bovine brain: a comparison with the dimeric G2 form of the electric organ.
Globular forms (G forms) of acetylcholinesterase (AChE) are formed by monomers, dimers and tetramers of the catalytic subunits (G1, G2 and G4). In this work the hydrophobic G2 and G4 AChE forms were purified to homogeneity from Discopyge electric organ and bovine caudate nucleus and studied from different points of view, including: velocity sedimentation, affinity to lectins and SDS-polyacrylamide gel electrophoresis under reducing and non-reducing conditions. The polypeptide composition of Discopyge electric organ G2 is similar to Torpedo, however the pattern of the brain G4 AChE is much complex. Under non-reducing conditions the catalytic subunit possesses a molecular weight of 65 kDa, however this value increases to 68 kDa after reduction, suggesting that intrachain-disulfide bonds are important in the folding of the catalytic subunits of the AChE. Also it was found that after mild proteolysis; the (125I)-TID-20 kDa fragment decreased its molecular weight to approximately 10 kDa with little loss of AChE activity. Finally, we suggest a model for the organization of the different domains of the hydrophobic anchor fragment of the G4 form.
Fuentes
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ME
Department of Cell Biology, Faculty of Biological Sciences, Catholic University of Chile, Santiago.
Inestrosa
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NC
D000110:Acetylcholinesterase / Q000032:analysis* / Q000302:isolation & purification; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002499:Centrifugation, Density Gradient; D004555:Electric Fish; D004557:Electric Organ / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K; D037102:Lectins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D012995:Solubility; D013696:Temperature; D014909:Wheat Germ Agglutinins
D037102:Lectins; D014909:Wheat Germ Agglutinins; D000110:Acetylcholinesterase; D012697:Serine Endopeptidases; D019286:Endopeptidase K