A BIOCHEMICAL AND CYTOLOGICAL ANALYSIS OF THE COMPLEX PERIPLASMIC FLAGELLA FROM SPIROCHAETA-AURANTIA
A BIOCHEMICAL AND CYTOLOGICAL ANALYSIS OF THE COMPLEX PERIPLASMIC FLAGELLA FROM SPIROCHAETA-AURANTIA
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DOI：
10.1128/jb.170.9.4023-4032.1988
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发表时间：
1988-09-01
影响因子：
3.2
通讯作者：
GREENBERG, EP
中科院分区：
文献类型：
--
作者：
BRAHAMSHA, B;GREENBERG, EP
分离出橙色螺旋毛菌的周质鞭毛，并发现其超微结构和生化复杂。一般来说，鞭毛丝直径为 18 至 20 nm，似乎由 11 至 13 nm 宽的内部区域和外层组成。钩基体区域由两个紧密并列的圆盘组成，圆盘通过杆连接到钩子。纯化鞭毛的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及产生钩和基体但不产生鞭毛丝的运动突变体的蛋白质印迹分析表明，丝由表观分子量为 37,500、34,000 和 31,500 的三种主要多肽（37.5K、34K 和 31.5K 多肽）和三种次要多肽组成。 36,000、33,000 和 32,000 表观分子量（36K、33K 和 32K 多肽）。纯化的钩基体制剂极大地富集了表观分子量在62,000至66,000范围内的三种多肽。使用特异于 37.5K 鞭毛蛋白的单克隆抗体以及与 36K、34K、33K、32K 和 31.5K 鞭毛蛋白共有的表位反应的免疫金标记实验表明，37.5K 主要多肽是外层的组成部分，而一种或多种其他多肽构成核心。
The periplasmic flagella of Spirochaeta aurantia were isolated and were found to be ultrastructurally and biochemically complex. Generally, flagellar filaments were 18 to 20 nm in diameter and appeared to consist of an 11 to 13-nm-wide inner region and an outer layer. The hook-basal body region consisted of two closely apposed disks connected to a hook by a rod. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified flagella together with a Western blot analysis of a motility mutant that produces hooks and basal bodies but not flagellar filaments revealed that the filaments were composed of three major polypeptides of 37,500, 34,000, and 31,500 apparent molecular weight (37.5K, 34K, and 31.5K polypeptides) and three minor polypeptides of 36,000, 33,000, and 32,000 apparent molecular weight (36K, 33K, and 32K polypeptides). Purified hook-basal body preparations were greatly enriched in three polypeptides in the range of 62,000 to 66,000 apparent molecular weight. Immunogold labeling experiments with a monoclonal antibody specific for the 37.5K flagellin and one reacts with an epitope common to the 36K, 34K, 33K, 32K, and 31.5K flagellins revealed that the 37.5K major polypeptide was a component of the outer layer, whereas one or more of the other polypeptides constituted the core.