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脊椎动物乙酰胆碱酯酶的多糖抗原决定簇及其抑制性单克隆抗体

Polysaccharide antigenic determinants against vertebrate acetylcholinesterases and their inhibitory monoclonal antibodies
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摘要 抗丁氏双鳍电鳐电器官乙酰胆碱酯酶(AChE)的抑制性单克隆抗体IH11(IgM),丁氏双鳍电鳐,石纹电线电器官,比目鱼肌肉,大鼠脑,牛脑,人脑,牛红细胞及人红细胞ACbE均以不同的亲和层析法提纯。用酶抗原免疫测定法及酶联免疫吸附测定法检测了1H11与上述脊椎动物的AChE的交叉免疫反应性.结果表明,1H11可与石纹电绥,比目鱼肌肉,大鼠脑、牛脑及人脑AChE有交叉免疫反应。显示了1H11所针对的抗原决定簇在进化中的高度保守性。1H11与人,牛红细胞AChE及人丁酰胆碱酯酶(BChE)无交叉免疫反应性。提示哺乳类脑AChE与红细胞AChE及人BChE有不同的抗原决定簇。1H11对电鳐及电鳗AChE活性产生明显抑制。但对其它几个脊椎动物的AChE活性无抑制.表明1H11所针对的抗原决定簇在电鳐AChE的三级结构中靠近其活性中心,而在其它脊椎动物的AChE中刚远离其活性中心.AChE阴离子部位抑制剂依酚氟铵不影响抗原抗体反应,表明1H11不结合在阴离子部位。N-糖苷酶F处理使ACbE去糖基化后。1H11不再能与AChE结合,表明至少1H11所识别的抗原决定簇的主要部分是N-多醣.抑制性单克隆抗体1H11对脑ACh? The monoclonal antibody 1H11(IgM)to Narcine timlei(Nt)acetylcholinesterase(AChE),and the AChEs from electric organs of Ntand Torpedo marmorata(Tm),flounder muscle,rat,bovine and human brains,bovine and human redblood cells(RBC)were purified by using different affin-ity chromatography.Enzyme antigen immunoassayrevealed that 1H11 exhibited strong crossimmunoreactivites with AChEs of Tm,flounder mus-cle,fat,bovine and human brains,manifesting the highconservation of the epitope recognized by 1H11 inevolution.No reacthity of 1H11 was observedn neitherWith human and bovine RBC AChEs,nor with humanbutyrylcholinesterase(BChE).Enzyme activity ofTm AChEs could be markedly inhibited by 1H11,whereas AChEs from other sources remaincd intact.Itprobably indicates that the 1H11-directed epitope lo-cates near the active center formed in the threedimentional structure in Tm AChEs,and is outsidethe area in the AChE molecules of other specics.Edrophonium did not influence the antigen-antibodyreaction, indicating that 1H11 does not bind at theanionic site of the active center.N-glycosidase F treat-ment of AChE caused 1H11 to lose the binding capaci-ty with the enzyme. This implies that the major part of1N11-directed epitope is carbohydrate in nature. Thehigh specificity of 1H11 could be used in distinguishingthe brain AChE from RBC AChE or serum BChE.
出处 《中国药理学与毒理学杂志》 CSCD 北大核心 1994年第3期213-217,共5页 Chinese Journal of Pharmacology and Toxicology
基金 国家自然科学基金
关键词 乙酰胆碱酯酶 单克隆抗体 脊椎动物 acetylcholinesterase butyrylcholi-nesterase antibodies monoclonal antigenicdeterminants deglycosylation
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  • 1辛颜彬,阎国珍,徐品芳,李凤珍,张翰,王家蓓,孙曼霁.抗乙酰胆碱酯酶单克隆抗体的产生[J].中国药理学与毒理学杂志,1989,3(1):68-72. 被引量:9
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