Purification and comparison of outer membrane protein P2 from Haemophilus influenzae type b isolates.
Purification and comparison of outer membrane protein P2 from Haemophilus influenzae type b isolates.
复制标题
b 型流感嗜血杆菌分离株外膜蛋白 P2 的纯化和比较。
DOI：
10.1172/jci111017
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Granoff,DM
中科院分区：
文献类型：
--
作者：
MunsonJr,RS;Shenep,JL;Barenkamp,SJ;Granoff,DM
根据梯度和改良Laemmli十二烷基硫酸钠-聚丙烯酰胺凝胶电泳系统上分离的主要外膜蛋白（OMP）图谱的差异，对B型流感嗜血杆菌分离株进行了细分。虽然已经观察到21种亚型，但86%的侵袭性分离株具有五种常见亚型之一，71%的分离株具有三种亚型之一。在具有两种最常见外膜亚型的分离株中，一种主要OMP的表观分子量为37，000。在具有另一种常见OMP亚型的分离株中，观察到表观分子量为36，500的交叉反应蛋白。这三种蛋白质都被命名为P2。将来自这些原型分离物的蛋白质P2用Zwittergent 3-14溶解并纯化至均一。基于氨基酸组成，溴化氰裂解产物，葡萄球菌V8蛋白酶，和胰凝乳蛋白酶肽图谱，P2蛋白从三个分离物已高度保守。用免疫印迹法检测了其中一株P2的兔抗抗体与另外两株P2的交叉反应。该抗体被动保护幼年大鼠免受由同源生物体引起的B型嗜血杆菌感染，但不抵抗具有异源36，500 mol wt P2蛋白的菌株的攻击。因此，尽管来自具有不同OMP亚型的分离株的P2蛋白密切相关，但保护性实验表明，与保护性抗体相互作用的细胞表面上的决定簇可能是菌株或亚型特异性的。
Haemophilus influenzae type b isolates have been subdivided based on differences in major outer membrane protein (OMP) profiles resolved on gradient and modified Laemmli sodium dodecyl sulfate-polyacrylamide gel electrophoresis systems. Although 21 subtypes have been observed, 86% of invasive isolates have one of five common subtypes and 71% of isolates have one of three subtypes. In isolates with two of the most common outer membrane subtypes, one major OMP has an apparent molecular weight of 37,000. In isolates with another common OMP subtype, a cross-reactive protein with an apparent molecular weight of 36,500 is observed. All three proteins have been designated P2. Protein P2 from these prototype isolates was solubilized with Zwittergent 3-14 and purified to homogeneity. Based on amino acid compositions, cyanogen bromide cleavage products, staphylococcal V8 protease, and chymotryptic peptide maps, the P2 protein from the three isolates has been highly conserved. Rabbit antibody prepared against P2 from one strain was cross-reactive with P2 isolated from the other two heterologous strains by Western blot. This antibody passively protected infant rats against type b Haemophilus infection caused by the homologous organism, but not against challenge by a strain with the heterologous 36,500 mol wt P2 protein. Thus, although the P2 protein from isolates with different OMP subtypes are closely related, the protection experiments suggest that determinants on the cell surface interacting with protective antibody may be strain or subtype specific.ImagesFIGURE 2FIGURE 3FIGURE 4FIGURE 5FIGURE 6FIGURE 7