Structural analysis of the variable major proteins of Borrelia hermsii.
Structural analysis of the variable major proteins of Borrelia hermsii.
复制标题
DOI：
10.1084/jem.158.6.2127
复制
发表时间：
1983-12-01
期刊：
影响因子：
--
通讯作者：
Judd RC
中科院分区：
文献类型：
--
作者：
Barbour AG;Barrera O;Judd RC
赫氏疏螺旋体在体内和体外经历自发抗原变异。血清型特异性与分子量可变蛋白家族之一（pI 蛋白）的表达相关。我们研究了赫氏芽孢杆菌 HS1 三种血清型：C、7 和 21 的 pI 蛋白以及分子量不变的 pII 蛋白的结构。所使用的技术是金黄色葡萄球菌 V8 蛋白酶生成的肽的一维 (1-D) 作图和 α-胰凝乳蛋白酶生成的肽的二维 (2-D) 作图。通过从染色的聚丙烯酰胺凝胶电泳图中切除多肽来分离pI和pII蛋白。一维肽模式通过本质上[14C]亮氨酸标记的蛋白质的荧光照相术或通过银染来可视化。在进行 2-D 作图之前，在氯胺-T 存在下用 125I 标记切下的凝胶片段中的多肽。我们还比较了 pI 蛋白、pI7 和 pI21 的表面暴露部分使用 1,3,4,6-四氯-3 α,6 α-二苯基甘脲（Iodogen）进行放射性碘化后的二维肽图。 I-D 和 2-D 肽图证明了以下内容：(a) 三种血清型的 pI 蛋白几乎没有共同的 V8 蛋白酶或胰凝乳蛋白酶生成的肽，并且 (b) 每种血清型的 pI 蛋白似乎相同。研究结果表明，pI 蛋白质变异性源自这些蛋白质氨基酸序列的广泛差异。
Borrelia hermsii undergoes spontaneous antigenic variation in vivo and in vitro. Serotype specificity is associated with expression of one of a family of molecular weight-variable proteins, the pI proteins. We studied the structure of the pI proteins as well as the molecular weight-invariable pII proteins of three serotypes of B. hermsii HS1: C, 7, and 21. The techniques used were one-dimensional (1-D) mapping of Staphylococcus aureus V8 protease-generated peptides and two- dimensional (2-D) mapping of alpha-chymotrypsin-generated peptides. The pI and pII proteins were isolated by excision of polypeptides from stained polyacrylamide gel electropherograms. The 1-D peptide patterns were visualized by fluorography of intrinsically [14C]leucine-labeled proteins or by silver stain. Before 2-D mapping, polypeptides in excised gel fragments were labeled with 125I in the presence of chloramine-T. We also compared the 2-D peptide maps of pI proteins, pI7 and pI21, after their surface-exposed portions were radioiodinated using 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril (Iodogen). The I-D and 2-D peptide maps demonstrated the following: (a) pI proteins of the three serotypes have few V8 protease- or chymotrypsin- generated peptides in common, and (b) pI proteins of each serotype appear to be identical. The findings suggest that pI protein variability derives from extensive differences in the amino acid sequences of these proteins.
登录
查看更多内容
影响因子：
3.1
作者：
JUDD, RC
通讯作者：
JUDD, RC
影响因子：
2.9
作者：
POEHLING, HM;NEUHOFF, V
通讯作者：
NEUHOFF, V
影响因子：
3.7
作者：
LAVER, WG;WEBSTER, RG
通讯作者：
WEBSTER, RG
影响因子：
2.9
作者：
MARKWELL, MAK
通讯作者：
MARKWELL, MAK
影响因子：
2.9
作者：
MARKWELL, MAK;FOX, CF
通讯作者：
FOX, CF