Freriksen A, Heinstra P W
Department of Plant Ecology and Evolutionary Biology, University of Utrecht, The Netherlands.
Biochem Genet. 1993 Oct;31(9-10):393-407.
Polyclonal antibodies raised against purified Drosophila alcohol dehydrogenase (ADH) were used in Western blot analyses to search for structurally and/or immunologically related proteins in prokaryotes and eukaryotes. No immunological-reactive protein was detected in a flesh fly, a locust, and butterflies. Immunological similarity with the 50-kDa PQQ-glucose dehydrogenase (GluDH)-B enzyme of Acinetobacter calcoaceticus was found, but the cross-reactivity apparently is dependent on the high hydrophilic character of this protein. Antibodies against PQQ-GluDH did not recognize Drosophila ADH. In five of seven species of the gram-positive soil bacteria actinomycetes tested, a protein approximately 28-30 kDa in subunit size was strongly recognized by alpha-DADH. It is probably not one of the two proteins with known homology to Drosophila ADH, viz., the actIII gene product and 20 beta-hydroxysteroid dehydrogenase. The protein is present in both the soluble and the pellet-membrane fraction of the cells. The protein has a late temporal expression in surface-grown cultures and, therefore, might be involved in secondary metabolism.
用针对纯化的果蝇乙醇脱氢酶(ADH)产生的多克隆抗体进行蛋白质免疫印迹分析,以寻找原核生物和真核生物中结构和/或免疫相关的蛋白质。在麻蝇、蝗虫和蝴蝶中未检测到免疫反应性蛋白。发现与乙酸钙不动杆菌的50 kDa吡咯喹啉醌葡萄糖脱氢酶(GluDH)-B酶存在免疫相似性,但这种交叉反应显然取决于该蛋白的高亲水性。抗PQQ-GluDH的抗体不能识别果蝇ADH。在测试的7种革兰氏阳性土壤细菌放线菌中的5种中,一种亚基大小约为28-30 kDa的蛋白质被α-DADH强烈识别。它可能不是与果蝇ADH具有已知同源性的两种蛋白质之一,即actIII基因产物和20β-羟基类固醇脱氢酶。该蛋白存在于细胞的可溶性部分和沉淀膜部分。该蛋白在表面生长的培养物中具有较晚的时间表达,因此可能参与次级代谢。