Pérez Carrillo Victor Hugo, Whittaker Jacob J, Wiedemann Christoph, Harder Jean-Martin, Lohr Theresa, Jamithireddy Anil K, Dajka Marina, Goretzki Benedikt, Joseph Benesh, Guskov Albert, Harmer Nicholas J, Holzgrabe Ulrike, Hellmich Ute A
Faculty of Chemistry and Earth Sciences, Institute of Organic Chemistry and Macromolecular Chemistry, Friedrich Schiller University Jena, 07743 Jena, Germany.
Groningen Institute for Biomolecular Sciences and Biotechnology, University of Groningen, 9747AG Groningen, The Netherlands.
J Med Chem. 2025 Mar 13;68(5):5926-5941. doi: 10.1021/acs.jmedchem.5c00134. Epub 2025 Feb 20.
Macrophage infectivity potentiator (MIP) proteins, found in pro- and eukaryotic pathogens, influence microbial virulence, host cell infection, pathogen replication, and dissemination. MIPs share an FKBP (FK506 binding protein)-like prolyl--isomerase domain, making them attractive targets for inhibitor development. We determined high-resolution crystal structures of and MIPs in complex with fluorinated pipecolic acid inhibitors. The inhibitor binding profiles in solution were compared across , , and MIPs using H, N, and F NMR spectroscopy. Demonstrating the versatility of fluorinated ligands for characterizing inhibitor complexes, F NMR spectroscopy identified differences in ligand binding dynamics across MIPs. EPR spectroscopy and SAXS further revealed inhibitor-induced global structural changes in homodimeric MIP. This study demonstrates the importance of integrating diverse methods to probe protein dynamics and provides a foundation for optimizing MIP-targeted inhibitors in this structurally conserved yet dynamically variable protein family.
巨噬细胞感染增强因子（MIP）蛋白存在于原核和真核病原体中，影响微生物毒力、宿主细胞感染、病原体复制和传播。MIPs具有一个类似FKBP（FK506结合蛋白）的脯氨酰异构酶结构域，这使得它们成为抑制剂开发的有吸引力的靶点。我们确定了与氟化哌啶酸抑制剂结合的和MIPs的高分辨率晶体结构。使用H、N和F NMR光谱比较了溶液中、和MIPs的抑制剂结合谱。F NMR光谱证明了氟化配体在表征抑制剂复合物方面的多功能性，确定了不同MIPs之间配体结合动力学的差异。EPR光谱和SAXS进一步揭示了抑制剂诱导的同二聚体MIP的整体结构变化。这项研究证明了整合多种方法来探测蛋白质动力学的重要性，并为优化这个结构保守但动态可变的蛋白质家族中针对MIP的抑制剂提供了基础。
