Pen J, Van Beeumen J, Beintema J J
Biochem J. 1986 Sep 15;238(3):691-9. doi: 10.1042/bj2380691.
Antibodies raised against esterase-4 and esterase-5 from Drosophila mojavensis were coupled to Protein A-Sepharose CL-4B to prepare high-efficiency immunomatrices used for their purification. Final purification was achieved by anion-exchange h.p.l.c., in the case of esterase-5 followed by gel-filtration h.p.l.c. The resultant esterase preparations were homogeneous, as judged by gel-filtration h.p.l.c., SDS/polyacrylamide-gel electrophoresis and non-denaturing gel electrophoresis. Esterase-4 and esterase-5 are the products of a duplicated gene. They are differently localized in the insect's body and expressed in different periods during development. Although both enzymes exhibit little immunological cross-reactivity, their amino acid compositions show few significant differences and their N-terminal sequences are largely identical, which clearly show their common origin.
针对来自莫哈韦果蝇的酯酶-4和酯酶-5产生的抗体与蛋白A-琼脂糖凝胶CL-4B偶联，以制备用于其纯化的高效免疫基质。酯酶-5的最终纯化通过阴离子交换高效液相色谱法实现，随后进行凝胶过滤高效液相色谱法。通过凝胶过滤高效液相色谱法、SDS/聚丙烯酰胺凝胶电泳和非变性凝胶电泳判断，所得酯酶制剂是均一的。酯酶-4和酯酶-5是一个重复基因的产物。它们在昆虫体内的定位不同，并且在发育的不同时期表达。尽管这两种酶几乎没有免疫交叉反应性，但它们的氨基酸组成几乎没有显著差异，并且它们的N端序列基本相同，这清楚地表明了它们的共同起源。
