Muensch H, Yoshida A, Altland K, Jensen W, Goedde H W
Am J Hum Genet. 1978 May;30(3):302-7.
Human plasma cholinesterase from five different genotypes -- E1U E1U, E1U E1A, E1A E1A, E1U E1S, E1A E1S, and E1U E1U C5+ -- was purified 8,000 fold from serum by a two-step procedure involving chromatography on DEAE-cellulose and preparative disc electrophoresis. The esterases were labeled with diisopropyl-1, 3-C14-fluorophosphate (DFP) aminoethylated, and digested by trypsin. The trytic digests were subjected to high voltage electrophoresis, and the radioactive peptides were detected by radioautography. Comparison of the peptides revealed different electrophoretic mobilities of the usual and atypical (dibucaine resistant) plasma cholinesterase peptides. The results are consistent with a structural abnormality of the active center in the variant enzyme. No difference was observed an the esteratic site of the enzyme with C5 component.
通过两步法从血清中纯化了来自五种不同基因型（E1U E1U、E1U E1A、E1A E1A、E1U E1S、E1A E1S和E1U E1U C5+）的人血浆胆碱酯酶，纯化倍数达8000倍，该两步法包括在DEAE - 纤维素上进行色谱分离和制备性圆盘电泳。用二异丙基 - 1,3 - C14 - 氟磷酸酯（DFP）对酯酶进行标记、氨乙基化，然后用胰蛋白酶消化。将胰蛋白酶消化产物进行高压电泳，通过放射自显影检测放射性肽段。肽段比较显示正常和非典型（耐丁卡因）血浆胆碱酯酶肽段具有不同的电泳迁移率。结果与变异酶活性中心的结构异常一致。在含有C5成分的酶的酯解部位未观察到差异。
