Subcellular localization of the Streptococcus mutans P1 protein C terminus.
Subcellular localization of the Streptococcus mutans P1 protein C terminus.
复制标题
变形链球菌 P1 蛋白 C 末端的亚细胞定位。
DOI：
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
G. Bowden
中科院分区：
文献类型：
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作者：
M. K. Homonylo;S. F. Lee;G. Bowden
为了确定变形链球菌P1蛋白C-末端锚的亚细胞位置，使用对定位在P1蛋白C-末端附近的表位具有特异性的单克隆抗体MAb 6- 8 C以及针对P1 C-末端144个氨基酸（P1 COOH）的多克隆抗体制剂，结合Western免疫印迹进行细胞包膜分级实验。P1蛋白在细胞壁中检测到，而在S.通过单克隆抗体对变形杆菌细胞的作用。相反，使用抗P1 COOH多克隆抗体在相同的细胞壁制备物中未检测到P1蛋白。然而，通过用变溶菌素处理从细胞壁释放的蛋白质含有被抗P1 COOH抗体识别的抗原，这表明被抗体识别的表位被细胞壁制剂中的肽聚糖掩蔽。当细胞壁用沸腾的三氯乙酸处理以溶解细胞壁相关的碳水化合物时，无论使用多克隆抗体还是单克隆抗体，在溶解的碳水化合物或剩余的肽聚糖中均不能检测到P1抗原。然而，当用变溶菌素处理肽聚糖时，P1抗原可以在变溶菌素溶解的组分中被MAb 6- 8 C检测到。总的来说，这些数据表明P1蛋白的C-末端144个氨基酸嵌入细胞壁内，并且仅与肽聚糖相关。此外，抗P1 COOH抗体仅在细胞壁的变溶菌素处理后识别P1抗原的能力表明这些C-末端144个氨基酸紧密嵌入肽聚糖链内。
To determine the subcellular location of the Streptococcus mutans P1 protein C-terminal anchor, cell envelope fractionation experiments were conducted in combination with Western immunoblotting, using monoclonal antibody MAb 6-8C specific for an epitope that maps near the C terminus of P1 protein and also a polyclonal antibody preparation directed against the P1 C-terminal 144 amino acids (P1COOH). P1 protein was detected in cell walls but not the membrane purified from S. mutans cells by the monoclonal antibody. In contrast, P1 protein was not detected in the same cell wall preparation using the anti-P1COOH polyclonal antibody. However, proteins released from the cell walls by treatment with mutanolysin contained antigen that was recognized by the anti-P1COOH antibody, suggesting that the epitopes recognized by the antibody were masked by peptidoglycan in the cell wall preparations. When cell walls were treated with boiling trichloroacetic acid to solubilize cell-wall-associated carbohydrate, P1 antigen could not be detected in either the solubilized carbohydrate, or in the remaining peptidoglycan, regardless of whether polyclonal or monoclonal antibody was used. However, when the peptidoglycan was treated with mutanolysin, P1 antigen could be detected in the mutanolysin solubilized fraction by MAb 6-8C. Collectively, these data suggest that the C-terminal 144 amino acids of the P1 protein are embedded within the cell wall, and associated exclusively with the peptidoglycan. Furthermore, the ability of the anti-P1COOH antibody to recognize P1 antigen only after mutanolysin treatment of cell walls suggests these C-terminal 144 amino acids are tightly intercalated within the peptidoglycan strands.
影响因子：
56.9
作者：
SCHNEEWIND, O;FOWLER, A;FAULL, KF
通讯作者：
FAULL, KF