Identification of amino acids important for binding of Clostridium perfringens epsilon toxin to host cells and to HAVCR1.
Identification of amino acids important for binding of Clostridium perfringens epsilon toxin to host cells and to HAVCR1.
复制标题
鉴定氨基酸对于结合灌注梭状芽胞杆菌与宿主细胞和HAVCR1的结合至关重要的氨基酸。
DOI：
10.1021/bi300690a
复制
发表时间：
2012-09-25
期刊：
影响因子：
2.9
通讯作者：
McClain MS
中科院分区：
文献类型：
--
作者：
Ivie SE;McClain MS
灌注梭状芽胞杆菌氏乳蛋白毒素属于孔形成毒素的类似气膜素家族，是已知的最有效的细菌毒素之一。伊普龙毒素会导致致命的肠毒素血症，山羊和可能的人类。证据表明，毒素与包括肝炎A病毒受体1（HAVCR1）的蛋白质受体结合，但尚未确定负责细胞结合的毒素区域。在本研究中，我们鉴定出对这种细胞相互作用很重要的Epsilon毒素中的氨基酸。位点特异性诱变用于研究表面可访问的芳香族氨基酸簇的作用，并在一系列的细胞培养分析中测试了纯化的突变蛋白，以评估细胞毒性活性和细胞结合。当添加到细胞中时，四种突变蛋白（ETX-Y29E，ETX-Y30E，ETX-Y36E和ETX-Y196E）不仅会严重损害其不仅杀死宿主细胞的能力，而且还具有渗透性质膜的能力。循环二分色谱和热稳定性研究表明，野生型和突变蛋白类似地折叠。其他实验表明，这些突变蛋白在与宿主细胞和HAVCR1的结合中有缺陷。这些数据表明，包括Y29，Y30，Y36和Y196在内的氨基酸基序对于Epsilon毒素与细胞和HAVCR1相互作用的能力很重要。
Clostridium perfringens epsilon toxin belongs to the aerolysin-like family of pore-forming toxins and is one of the most potent bacterial toxins known. The epsilon toxin causes fatal enterotoxemia in sheep, goats, and possibly humans. Evidence indicates that the toxin binds to protein receptors including hepatitis A virus cellular receptor 1 (HAVCR1), but the region of the toxin responsible for cell binding has not been identified. In the present study, we identify amino acids within the epsilon toxin important for this cell interaction. Site-specific mutagenesis was used to investigate the role of a surface-accessible cluster of aromatic amino acids, and purified mutant proteins were tested in a series of cell-culture assays to assess cytotoxic activity and cell binding. When added to cells, four mutant proteins (Etx-Y29E, Etx-Y30E, Etx-Y36E and Etx-Y196E) were severely impaired in their ability to not only kill host cells, but also in their ability to permeabilize the plasma membrane. Circular dichroism spectroscopy and thermal stability studies revealed that the wild-type and mutant proteins were similarly folded. Additional experiments revealed that these mutant proteins were defective in binding to host cells and to HAVCR1. These data indicate that an amino acid motif including Y29, Y30, Y36, and Y196 is important for the ability of epsilon toxin to interact with cells and HAVCR1.
登录
查看更多内容
影响因子：
3.7
作者：
Goldstein J;Morris WE;Loidl CF;Tironi-Farinati C;McClane BA;Uzal FA;Fernandez Miyakawa ME
通讯作者：
Fernandez Miyakawa ME
影响因子：
4.2
作者：
Li YM;Hall WA
通讯作者：
Hall WA
DOI：
10.1002/path.1700710206
发表时间：
1956-01-01
期刊：
JOURNAL OF PATHOLOGY AND BACTERIOLOGY
影响因子：
--
作者：
BULLEN, JJ;BATTY, I
通讯作者：
BATTY, I
影响因子：
2.9
作者：
COMPTON, LA;JOHNSON, WC
通讯作者：
JOHNSON, WC
影响因子：
4.8
作者：
MacKenzie, CR;Hirama, T;Buckley, JT
通讯作者：
Buckley, JT