PT - JOURNAL ARTICLE AU - C. Wiedemann AU - J.J. Whittaker AU - V.H. Pérez Carrillo AU - B. Goretzki AU - M. Dajka AU - F. Tebbe AU - J.-M. Harder AU - P. Krajczy AU - B. Joseph AU - F. Hausch AU - A. Guskov AU - U.A. Hellmich TI - <em>Legionella pneumophila</em> macrophage infectivity potentiator protein appendage domains modulate protein dynamics and inhibitor binding AID - 10.1101/2023.04.24.538046 DP - 2023 Jan 01 TA - bioRxiv PG - 2023.04.24.538046 4099 - http://biorxiv.org/content/early/2023/04/26/2023.04.24.538046.short 4100 - http://biorxiv.org/content/early/2023/04/26/2023.04.24.538046.full AB - Macrophage infectivity potentiator (MIP) proteins are widespread in human pathogens including Legionella pneumophila, the causative agent of Legionnaires’ disease and protozoans such as Trypanosoma cruzi. All MIP proteins contain a FKBP (FK506 binding protein)-like prolyl-cis/trans-isomerase domain that hence presents an attractive drug target. Some MIPs such as the Legionella protein (LpMIP) have additional appendage domains of mostly unknown function. In full-length, homodimeric LpMIP, the N-terminal dimerization domain is linked to the FKBP-like domain via a long, free-standing stalk helix. Combining X-ray crystallography, NMR and EPR spectroscopy and SAXS, we elucidated the importance of the stalk helix for protein dynamics and inhibitor binding to the FKBP-like domain and bidirectional crosstalk between the different protein regions. The first comparison of a microbial MIP and a human FKBP in complex with the same synthetic inhibitor was made possible by high-resolution structures of LpMIP with a [4.3.1]-aza-bicyclic sulfonamide and provides a basis for designing pathogen-selective inhibitors. Through stereospecific methylation, the affinity of inhibitors to to L. pneumophila and T. cruzi MIP was greatly improved. The resulting X-ray inhibitor-complex structures of LpMIP and TcMIP at 1.49 and 1.34 Å, respectively, provide a starting point for developing potent inhibitors against MIPs from multiple pathogenic microorganisms.Competing Interest StatementThe authors have declared no competing interest.