Reyes A E, Perez D R, Alvarez A, Garrido J, Gentry M K, Doctor B P, Inestrosa N C
Departamento de Biología Celular y Molecular, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
Biochem Biophys Res Commun. 1997 Mar 27;232(3):652-5. doi: 10.1006/bbrc.1997.6357.
A monoclonal antibody (mAb) 25B1 directed against fetal bovine-serum acetylcholinesterase (FBS AChE) was used to examine the ability of the cholinergic enzyme to promote the assembly of amyloid-beta peptides (A beta) into Alzheimerś fibrils. This mAb binds to the peripheral anionic site of the enzyme and allosterically inhibits catalytic activity of FBS AChE. Several techniques, including thioflavine-T fluorescence, turbidity, and negative-staining at the electron microscopy level, were used to assess amyloid formation. Inhibition of amyloid formation was dependent on the molar ratio AChE:mAb 25B1, and at least 50% of the inhibition of the AChE promoting effect occurs at a molar ratio similar to that required for inhibition of the esterase activity. Our results suggest that mAb 25B1 inhibits the promotion of the amyloid fibril formation triggered by AChE by affecting the lag period of the A beta aggregation process.
一种针对胎牛血清乙酰胆碱酯酶（FBS AChE）的单克隆抗体（mAb）25B1被用于检测胆碱能酶促进β淀粉样肽（Aβ）组装成阿尔茨海默病纤维的能力。这种单克隆抗体与该酶的外周阴离子位点结合，并变构抑制FBS AChE的催化活性。使用了几种技术，包括硫黄素-T荧光、浊度以及电子显微镜水平的负染色，来评估淀粉样蛋白的形成。淀粉样蛋白形成的抑制取决于AChE与mAb 25B1的摩尔比，并且至少50%的AChE促进作用的抑制发生在与抑制酯酶活性所需摩尔比相似的情况下。我们的结果表明，mAb 25B1通过影响Aβ聚集过程的延迟期来抑制由AChE触发的淀粉样纤维形成。
