A Novel Cholesterol-Dependent Cytolysin Receptor

描述（由申请方提供）：胆固醇依赖性溶细胞素（CDC）是由超过20种革兰氏阳性病原菌产生的相关细菌毒素的大家族。几种CDC已被证明有助于这些细菌物种的致病机制。CDC作为可溶性蛋白质产生，其在含胆固醇的膜的表面上寡聚成大孔形成复合物。膜胆固醇的存在是CDC孔形成所需的，并且是CDC的标志。三十多年来，CDC的受体通常被认为是胆固醇，但最近的研究表明，这是CDC的前孔到孔转化所必需的，这是膜结合下游的事件。因此，CDC使用胆固醇作为其受体的长期教条似乎不太可信。我们现在已经确定CDC家族的至少一个成员，来自中间链球菌的中间溶素（ILY），利用GPI锚定的人CD 59（hCD 59）作为其受体。基于初步研究，我们假设1）ILY结构域4含有与hCD 59结合并对hCD 59具有特异性所需的残基，2）由人补体蛋白C8和C9识别的hCD 59的区域和残基也包含ILY的hCD 59结合位点，和3）ILY在孔形成之前从其受体脱离。该提议旨在研究ILY与hCD 59的相互作用，并绘制出ILY和hCD 59的结构域，这些结构域参与这种配体-受体相互作用。还将在细胞溶解机制的各个阶段检查ILY与hCD 59的相互作用，以确定毒素是否在其孔复合物的组装期间从受体脱离。这些假设将在三个具体目标中得到解决，这三个具体目标将建立hCD 59和ILY上结合位点的位置，并将确定ILY是否在孔形成复合物组装期间从受体脱离。这些研究的结果将提供一个新的和重要的方面的CDC以前无法用于这些毒素的研究调查的基础。CDC受体的发现和研究将对CDC的研究产生重大影响，并将改变我们对它们在由各种革兰氏阳性病原体引起的大量疾病中的作用的看法。

DESCRIPTION (provided by applicant): The cholesterol-dependent cytolysins (CDCs) are a large family of related bacterial toxins produced by over 20 species of Gram-positive pathogenic bacteria. Several CDCs have been shown to contribute to the pathogenic mechanisms of these bacterial species. The CDCs are produced as soluble proteins that oligomerize into large pore-forming complexes on the surface of cholesterol-containing membranes. The presence of membrane cholesterol is required for the CDC pore-formation and is a hallmark of the CDCs. For over three decades the receptor for the CDCs was generally accepted as cholesterol but recent studies show that it is necessary for the prepore to pore conversion of the CDCs, an event that is downstream of membrane binding. Therefore, the longstanding dogma that the CDCs use cholesterol as their receptor appears less plausible. We have now determined that at least one member of the CDC family, intermedilysin (ILY) from Streptococcus intermedius, utilizes the GPI-anchored human CD59 (hCD59) as its receptor. Based on preliminary studies we hypothesize that 1) ILY domain 4 contains the residues required for binding to and specificity for hCD59, 2) the region and residues of hCD59 recognized by human complement proteins C8 and C9 also comprises the hCD59 binding site for ILY, and 3) ILY disengages from its receptor prior to pore formation. This proposal is designed to examine the interaction of ILY with hCD59 and to map out the structural domains of both ILY and hCD59 that are involved in this ligand-receptor interaction. The interaction of ILY with hCD59 will also be examined at various stages of the cytolytic mechanism to determine if the toxin disengages from receptor during the assembly of its pore complex. These hypotheses will be addressed in three specific aims that will establish the location of the binding sites on both hCD59 and ILY and will determine whether ILY undocks from receptor during the assembly of the pore-forming complex. The results of these studies will provide a basis for investigations of a new and significant aspect of the CDCs previously unavailable to the study of these toxins. The discovery and study of a CDC receptor will have a significant impact on the study of CDCs and will alter our perception of their role in a large number of diseases caused by a wide variety of Gram-positive pathogens.