Samaddar Manalee, Sun Chen, Gallina Nicholas L F, Irizarry-Tardi Nicole, Liu Dongqi, Tenguria Shivendra, Drolia Rishi, Jain Anika, Kihara Daisuke, Jiang Wen, Kim Kee-Hong, Cox Abigail, Ristroph Kurt D, Knipp Gregory T, Noinaj Nicholas, Bhunia Arun K
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN, USA.
Purdue Institute of Inflammation, Immunology, and Infectious Disease, Purdue University, West Lafayette, IN, USA.
Res Sq. 2025 May 14:rs.3.rs-6474377. doi: 10.21203/rs.3.rs-6474377/v1.
During infection, adhesion protein (LAP), a housekeeping enzyme, acts as a tight junction modulator (TJM) through interaction with Hsp60 to facilitate translocation across the intestinal epithelial barrier. Here, we used purified LAP as a potential TJM to overcome the limiting and variable effects observed by other agents in the class. We structurally determined the LAP interaction alone and in complex with Hsp60 utilizing cryo-EM and computational analysis. LAP structure resolved at 2.83 Å, forms multimeric interlocking dimers and tetramers, and the N-domain interacts with Hsp60, while the C-domain bridges bacterial surface receptor InlB. The structural studies complement LAP-mediated cyclic peptide drugs (vancomycin and desmopressin) absorption across the intestinal barrier in a mouse model without inducing inflammation or adverse effects on the TJ architecture. This study demonstrates that the LAP-Hsp60 complex is the basis for downstream utilization of LAP or peptide mimetics as promising TJM for improved peroral biologics delivery.
在感染过程中，粘附蛋白（LAP）作为一种管家酶，通过与热休克蛋白60（Hsp60）相互作用，充当紧密连接调节剂（TJM），以促进其穿过肠道上皮屏障的转运。在此，我们使用纯化的LAP作为一种潜在的TJM，以克服该类中其他药物所观察到的局限性和可变效应。我们利用冷冻电镜和计算分析，在结构上确定了LAP单独以及与Hsp60形成复合物时的相互作用。LAP结构在2.83 Å分辨率下解析，形成多聚体互锁二聚体和四聚体，其N结构域与Hsp60相互作用，而C结构域则连接细菌表面受体InlB。这些结构研究补充了LAP介导的环肽药物（万古霉素和去氨加压素）在小鼠模型中穿过肠道屏障的吸收情况，且不会诱导炎症或对紧密连接结构产生不良影响。这项研究表明，LAP-Hsp60复合物是将LAP或肽模拟物作为有前景的TJM用于改善口服生物制剂递送的下游应用的基础。
