Purification and Characterization of Axial Filaments from Treponema phagedenis Biotype reiterii (the Reiter Treponeme)
Purification and Characterization of Axial Filaments from Treponema phagedenis Biotype reiterii (the Reiter Treponeme)
复制标题
噬菌体密螺旋体生物型 reiterii（Reiter Treponeme）轴向丝的纯化和表征
DOI：
10.1128/jb.120.3.1434-1442.1974
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发表时间：
1974
影响因子：
3.2
通讯作者：
D. Allis
中科院分区：
文献类型：
--
作者：
M. Bharier;D. Allis
轴丝已被纯化从梅毒螺旋体phagedenis生物型reiterii（赖特密螺旋体）和部分化学特征。该制剂主要由蛋白质组成，但也含有少量的己糖（3%）。纤维在酸、碱、尿素、胍和各种洗涤剂中解离成亚基。氨基酸分析表明，整体相似的其他螺旋体轴丝和细菌鞭毛。在pH 4.3（在4 M尿素和10 - 3 M乙二胺四乙酸）和pH 12的丙烯酰胺凝胶电泳时，解离的细丝迁移作为一个单一的带，但在十二烷基硫酸钠凝胶中，三个带在各种各样的条件下获得。这些条带中的两条迁移得非常接近，分子量为33，000 ± 500。另一条带的分子量为36，500 ± 500。通过丹磺酰氯方法分析轴向丝产生甲硫氨酸和谷氨酸作为氨基末端基团。在7 M盐酸胍中对解离的轴向丝进行沉降平衡测量，得到log C对μ 2的曲线，该曲线随所使用的速度和初始蛋白质浓度而变化。从这些图计算的分子量值与其中轴向丝亚基相对于分子量在32，000至36，000的近似范围内是不均匀的模型一致。
Axial filaments have been purified from Treponema phagedenis biotype reiterii (the Reiter treponeme) and partially characterized chemically. The preparations consist largely of protein but also contain small amounts of hexose (3%). Filaments dissociate to subunits in acid, alkali, urea, guanidine, and various detergents. Amino acid analyses show an overall resemblance to other spirochetal axial filaments and to bacterial flagella. Dissociated filaments migrate as a single band upon acrylamide gel electrophoresis at pH 4.3 (in 4 M urea and 10 3 M ethylenediaminetetraacetate) and at pH 12, but in sodium dodecyl sulfate gels, three bands are obtained under a wide variety of conditions. Two of these bands migrate very close together, with molecular weights of 33,000 ± 500. The other band has a molecular weight of 36,500 ± 500. Analysis of axial filaments by the dansyl chloride method yields both methionine and glutamic acid as amino terminal end groups. Sedimentation equilibrium measurements on dissociated axial filaments in 7 M guanidine hydrochloride yield plots of log C against ϰ2 which vary with the speed and initial protein concentration used. Molecular weight values calculated from these plots are consistent with a model in which axial filament subunits are heterogeneous with respect to molecular weight in the approximate range of 32,000 to 36,000.