EMPIGEN BB - A USEFUL DETERGENT FOR SOLUBILIZATION AND BIOCHEMICAL-ANALYSIS OF KERATINS
EMPIGEN BB - A USEFUL DETERGENT FOR SOLUBILIZATION AND BIOCHEMICAL-ANALYSIS OF KERATINS
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DOI：
10.1006/bbrc.1995.1051
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发表时间：
1995-01-05
影响因子：
3.1
通讯作者：
OMARY, MB
中科院分区：
文献类型：
--
作者：
LOWTHERT, LA;KU, NO;OMARY, MB
中间丝（IF）蛋白质组成了一些已知的最不溶性的蛋白质，在IF蛋白质家族中，角蛋白是最不可溶的。我们比较了非离子型、阳离子型、混合型非离子型和阴离子型以及两性离子型洗涤剂在溶解来自表达重组人角蛋白的昆虫细胞以及来自人结肠细胞系和正常角质形成细胞的角蛋白方面的效率。阳离子洗涤剂十六烷基三甲基溴化铵在其有效溶解角蛋白的能力方面与两性离子洗涤剂Empigen BB相似，但后者的洗涤剂在保持抗体反应性并允许角蛋白的免疫沉淀方面是上级的。虽然Nonidet-P40部分溶解角蛋白，Empigen BB溶解了大量Nonidet-P40不溶解的角蛋白。就波形蛋白而言，洗涤剂之间溶解效率的差异并不像角蛋白那样显着。我们的研究结果表明，Empigen BB溶解了大量的表皮和腺角蛋白，同时保留抗原性。这种去污剂应该证明对这些蛋白质进行生物化学和分子研究是有用的，并且可能对其他IF蛋白质同样有益。(C)出版社：Academic Press
Intermediate filament (IF) proteins make up some of the most insoluble proteins known, and within the IF protein family, keratins are the least soluble. We compared the efficiency of nonionic, cationic, mixed nonionic and anionic, and zwitterionic detergents in solubilizing keratins from insect cells that express recombinant human keratins and from human colonic cell lines and normal keratinocytes. The cationic detergent cetyl trimethylammonium bromide was similar to the zwitterionic detergent Empigen BB in its ability to efficiently solubilize keratins, but the latter detergent was superior in that it maintained antibody reactivity and allowed for immunoprecipitation of the keratins. Although Nonidet-P40 partially solubilizes keratins, Empigen BB solubilizes a significant amount of keratins not solubilized by Nonidet-P40. In the case of vimentin, differences in solubilization efficiency among the detergents was not as dramatic as with keratins. Our results show that Empigen BB solubilizes a significant amount of epidermal and glandular keratins while preserving antigenicity. This detergent should prove useful for carrying out biochemical and molecular studies on these proteins and may be similarly beneficial for other IF proteins. (C) 1995 Academic Press, Inc.