Schryvers A B
Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
J Med Microbiol. 1989 Jun;29(2):121-30. doi: 10.1099/00222615-29-2-121.
An affinity procedure with purified, biotinylated human transferrin and streptavidin-agarose was used to identify the transferrin-binding proteins in strains of Haemophilus influenzae. Proteins of 58 and 98 Kda were isolated from total membranes prepared from iron-deficient but not iron-sufficient H. influenzae KC548 cells. The 58-Kda protein was capable of binding human transferrin after sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE) and electroblotting. Isolation of transferrin-binding proteins from type-b and non-typable H. influenzae strains demonstrated some variability in the size of the higher mol. wt protein (94-106 Kda) and in ease of elution of the smaller protein from the affinity resin. Use of purified, biotinylated human lactoferrin in the affinity isolation procedure with membranes from a strain expressing lactoferrin-binding activity resulted in isolation of proteins of 105 and 106 Kda distinct from the transferrin-binding proteins.
采用纯化的生物素化人转铁蛋白和链霉亲和素琼脂糖亲和程序，来鉴定流感嗜血杆菌菌株中的转铁蛋白结合蛋白。从缺铁而非富铁的流感嗜血杆菌KC548细胞制备的总膜中分离出了58 kDa和98 kDa的蛋白质。58 kDa的蛋白质在十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶电泳（PAGE）和电印迹后能够结合人转铁蛋白。从b型和不可分型流感嗜血杆菌菌株中分离转铁蛋白结合蛋白表明，较高分子量蛋白（94 - 106 kDa）的大小以及较小蛋白从亲和树脂上洗脱的难易程度存在一些差异。在对具有乳铁蛋白结合活性的菌株的膜进行亲和分离程序时，使用纯化的生物素化人乳铁蛋白，结果分离出了与转铁蛋白结合蛋白不同的105 kDa和106 kDa的蛋白质。
