Nikolau B J, Wurtele E S, Stumpf P K
Anal Biochem. 1985 Sep;149(2):448-53. doi: 10.1016/0003-2697(85)90596-2.
A procedure to detect biotinyl proteins after fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was developed. Proteins were immobilized on nitrocellulose and biotin-containing proteins were detected by probing with 125I-streptavidin. Using this procedure a small survey of biotinyl protein in plants was undertaken. In total four biotin-containing proteins were detected in higher plants of molecular weights 62,000, 50,000, 34,000, and 31,000. These biotinyl proteins were not ubiquitous in the plants surveyed. In the cyanobacterium Anabeana variabilis, a single biotin-containing protein of 21,000 Da was detected. In isolated spinach chloroplasts, the two biotinyl proteins detected were soluble. The results are discussed in relation to acetyl-CoA carboxylase.
开发了一种在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分级分离后检测生物素化蛋白的方法。将蛋白质固定在硝酸纤维素膜上,并用125I-链霉亲和素进行探测以检测含生物素的蛋白质。利用该方法对植物中的生物素化蛋白进行了小规模调查。在分子量为62000、50000、34000和31000的高等植物中总共检测到四种含生物素的蛋白质。这些生物素化蛋白在所调查的植物中并非普遍存在。在蓝藻多变鱼腥藻中,检测到一种分子量为21000 Da的含生物素蛋白。在分离的菠菜叶绿体中,检测到的两种生物素化蛋白是可溶的。结合乙酰辅酶A羧化酶对结果进行了讨论。