Whole Cell Cross-Linking to Discover Host-Microbe Protein Cognate Receptor/Ligand Pairs.
Whole Cell Cross-Linking to Discover Host-Microbe Protein Cognate Receptor/Ligand Pairs.
复制标题
DOI：
10.3389/fmicb.2018.01585
复制
发表时间：
2018
影响因子：
5.2
通讯作者：
Shah J
中科院分区：
文献类型：
--
作者：
Weimer BC;Chen P;Desai PT;Chen D;Shah J
细菌表面配体在结合过程中介导与宿主细胞的相互作用，这决定了宿主-微生物结合的具体结果。这种结合始于宿主细胞上的受体与微生物细胞上的配体结合，形成伙伴关系，从而在两个细胞中启动应答。缺乏确定具体同源伙伴关系的方法。确定宿主和微生物表面之间的这些分子相互作用是困难的，但对于定义微生物组结合过程中触发的重要生物学事件至关重要，对于定义来自宿主膜的导致病理或共生结合的启动信号至关重要。在这项研究中，我们设计了一种方法，利用整个细胞的共价交联策略来发现同源宿主-微生物受体/配体对。当相互作用的分子在9-12 Å范围内时，蛋白质/蛋白质交联发生，允许鉴定来自宿主和微生物的特定蛋白质对，这些蛋白质对定义了相互作用过程中的分子相互作用。为了验证该方法，研究人员检查了三种不同的细菌，它们具有三种已知的蛋白质/蛋白质伙伴关系。确切的相互作用得到了证实，并导致发现了额外的伙伴关系，这些伙伴关系不被认为是同源伙伴，但以前曾报道过与细菌相互作用有关。此外，通过阻断假定的宿主受体和删除细菌配体，发现了三个未知的受体/配体伙伴，并通过体外感染试验进行了验证。随后，肠沙门氏菌感染。鼠伤寒杆菌与分化的结肠上皮细胞（caco-2）交联，发现四种以前未知的宿主受体与三种以前未定义的沙门氏菌宿主配体结合。这种方法导致了先前未知的和生物学上重要的宿主/微生物关联分子的先验发现，这些分子被偶然报道介导细菌入侵。全细胞交联方法有望发现可能的靶标来调节微生物组与宿主的相互作用，这在感染和共生中都很重要，两者都启动宿主反应。
Bacterial surface ligands mediate interactions with the host cell during association that determines the specific outcome for the host–microbe association. The association begins with receptors on the host cell binding ligands on the microbial cell to form a partnership that initiates responses in both cells. Methods to determine the specific cognate partnerships are lacking. Determining these molecular interactions between the host and microbial surfaces are difficult, yet crucial in defining biologically important events that are triggered during association of the microbiome, and critical in defining the initiating signal from the host membrane that results in pathology or commensal association. In this study, we designed an approach to discover cognate host–microbe receptor/ligand pairs using a covalent cross-linking strategy with whole cells. Protein/protein cross-linking occurred when the interacting molecules were within 9–12 Å, allowing for identification of specific pairs of proteins from the host and microbe that define the molecular interaction during association. To validate the method three different bacteria with three previously known protein/protein partnerships were examined. The exact interactions were confirmed and led to discovery of additional partnerships that were not recognized as cognate partners, but were previously reported to be involved in bacterial interactions. Additionally, three unknown receptor/ligand partners were discovered and validated with in vitro infection assays by blocking the putative host receptor and deleting the bacterial ligand. Subsequently, Salmonella enterica sv. Typhimurium was cross-linked to differentiated colonic epithelial cells (caco-2) to discover four previously unknown host receptors bound to three previously undefined host ligands for Salmonella. This approach resulted in a priori discovery of previously unknown and biologically important molecules for host/microbe association that were casually reported to mediate bacterial invasion. The whole cell cross-linking approach promises to enable discovery of possible targets to modulate interaction of the microbiome with the host that are important in infection and commensalism, both of with initiate a host response.
登录
查看更多内容
自然界中过度感染细菌菌株出现的种内变化。
影响因子：
6.7
作者：
Heithoff DM;Shimp WR;House JK;Xie Y;Weimer BC;Sinsheimer RL;Mahan MJ
通讯作者：
Mahan MJ
DOI：
10.1128/genomea.00341-17
发表时间：
2017-05-01
期刊：
GENOME ANNOUNCEMENTS
影响因子：
--
作者：
Arabyan, Narine;Weis, Allison M.;Weimer, Bart C.
通讯作者：
Weimer, Bart C.
影响因子：
56.9
作者：
Daley, DO;Rapp, M;von Heijne, G
通讯作者：
von Heijne, G
影响因子：
3.5
作者：
Altermann, E;Buck, BL;Klaenhammer, TR
通讯作者：
Klaenhammer, TR
影响因子：
64.8
作者：
Chicurel, ME;Singer, RH;Ingber, DE
通讯作者：
Ingber, DE