人脑乙酰胆碱酯酶的分离纯化及特性

人脑纹状体及部分丘脑AChE在低盐缓冲液中以1％Triton X-100溶脱后,经Con A、短臂及长臂凝胶配基亲和层析纯化在还原条件下SDS-PAGE呈一主带,纯化的AChE比活性可达3384U/mg,产率20％,其单体分子量66 kD,聚丙烯酰胺凝胶电泳显示有单体及二、四、六、八聚体存在,等电点在pH5.3—6.0之间,人脑AChE活性大部分能用Con A亲和层析法回收说明为糖蛋白,纯化的人脑AChE与抗电鳐电器官AChE单克隆抗体3F3无抗原抗体反应,而与IH11及2G 8有弱的交叉反应,以Con A及短臂凝胶配基亲和层析纯化的人小脑AChE为抗原免疫Balb/c小鼠产生的抗血清与电鳐电器官AChE有较强的免疫交叉反应而与人红细胞膜AChE反应较弱,纯化的人脑AChE经还原羧甲基化及TPCK处理的胰蛋白酶切割后,再用反相HPLC分离,显示几个大肽峰及许多小肽峰,几个大肽峰以ELISA法测定有抗原性。     

我国的新药研究与开发

我国的新药研究与开发孙曼霁，周廷冲（军事医学科学院，毒物药物研究所，基础医学研究所，北京１００８５０）医药是人们战胜疾病的重要手段，它保障了人类的生存和繁衍，在人类漫长的发展史中起过并将继续发挥重要的作用。随着生活水平的提高，人们对药物的需求也日益增...     

Purification and Properties of Acetylcholinesterase From Human Brain

Acetylcholinesterase from human caudate nucleus and partial thalamus was purified by using Con A-Sepharose, short-arm and long-armligand Sepharose affinity chromatographies. SDS-PAGE of the purified AChE under the reduced condition showed one main band, corresponding to a molecular weight of 66 kD. The purified AChE witha specific activity of 3384 U/mg protein represented 20% activity of the homogenate supernatant. Analysis of purified AChE by gradient slab PAGE and DISC-PAGE with activity staining revealed the existence of monomer, dimer, tetramer, hexamer and octomer of the enzyme. The isoelectric point of AChE ranged between pH 5.6 and 6.0. Con A-Sepharose affinity chromatography retained most of the applied AChE activity implying that the enzyme is a kind of glycoprotein. The isolated human brain AChE had no cross-immunoreactivity with 3F3 and weak cross-immunoreactivity with 2G8 and 1H11 anti-Torpedo AChE antibodies. Balb/c mice were immunized with human cerebellum AChE purified with Con A and shor