MIF intersubunit disulfide mutant antagonist supports activation of CD74 by endogenous MIF trimer at physiologic concentrations
Chengpeng Fan,Deepa Rajasekaran,Mansoor Ali Syed,Lin Leng,J. Patrick Loria,Vineet Bhandari,Richard Bucala,Elias Lolis +7 more
TLDR
NMR and size-exclusion chromatography with light scattering reveal that N110C can form a higher-order oligomer in equilibrium with a single locked trimer and the X-ray structure confirms a local conformational change that disrupts the subunit interface and results in global changes responsible for the oligomeric form.Abstract:
Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine. In addition to its known receptor-mediated biological activities, MIF possesses a catalytic site of unknown function between subunits of a homotrimer. Each subunit contributes three β-strands to adjacent subunits to form a core seven-stranded β-sheet for each monomer. MIF monomers, dimers, or trimers have been reported, but the active form that binds and activates the MIF receptor (CD74) is still a matter of debate. A cysteine mutant (N110C) that covalently locks MIF into a trimer by forming a disulfide with Cys-80 of an adjacent subunit is used to study this issue. Partial catalytic activity and receptor binding to CD74 are retained by N110C (locked trimer), but there is no cellular signaling. Wild-type MIF-induced cellular signaling, in vivo lung neutrophil accumulation, and alveolar permeability are inhibited with a fivefold excess of N110C. NMR and size-exclusion chromatography with light scattering reveal that N110C can form a higher-order oligomer in equilibrium with a single locked trimer. The X-ray structure confirms a local conformational change that disrupts the subunit interface and results in global changes responsible for the oligomeric form. The structure also confirms these changes are consistent for the partial catalytic and receptor binding activities. The absence of any potential monomer and the retention of partial catalytic and receptor binding activities despite changes in conformation (and dynamics) in the mutant support an endogenous MIF trimer that binds and activates CD74 at nanomolar concentrations. This conclusion has implications for therapeutic development.read more
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Iterative model-building, structure refinement, and density modification with the PHENIX AutoBuild Wizard
TL;DR: The highly automated PHENIX AutoBuild wizard is described, which can be applied equally well to phases derived from isomorphous/anomalous and molecular-replacement methods.
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Cystic fibrosis sputum DNA has NETosis characteristics and neutrophil extracellular trap release is regulated by macrophage migration-inhibitory factor.
Markryan Dwyer,Qiang Shan,Samantha D'Ortona,Rie Maurer,Robert A. Mitchell,Hanne Olesen,Steffen Thiel,Johannes Huebner,Mihaela Gadjeva +8 more
TL;DR: It is suggested that targeting MIF by small molecular inhibitors might reduce the presence of extracellular DNA and serve as an adjunct to the use of antimicrobial drugs that could ultimately reduce bacterial fitness in the lungs during the later stages of CF disease.
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MIF, a controversial cytokine: a review of structural features, challenges, and opportunities for drug development
TL;DR: The field of anti-MIF therapeutics would benefit from capitalizing on the protein’s multiple assets while acknowledging their flaws, and should focus on developing more robust assays for MIF bioactivity that can be used for second-pass screening and specificity studies.
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Giant tortoise genomes provide insights into longevity and age-related disease
Víctor Quesada,Sandra Freitas-Rodríguez,Joshua M. Miller,José G. Pérez-Silva,Zi-Feng Jiang,Washington Tapia,Olaya Santiago-Fernández,Diana Campos-Iglesias,Lukas F. K. Kuderna,Maud C. Quinzin,Miguel G. Álvarez,Dido Carrero,Luciano B. Beheregaray,James P. Gibbs,Ylenia Chiari,Scott Glaberman,Claudio Ciofi,Miguel Araujo-Voces,Pablo Mayoral,Javier R. Arango,Isaac Tamargo-Gómez,David Roiz-Valle,María Pascual-Torner,Benjamin R. Evans,Danielle L. Edwards,Ryan C. Garrick,Michael A. Russello,Nikos Poulakakis,Nikos Poulakakis,Stephen J. Gaughran,Danny Rueda,Gabriel Bretones,Tomas Marques-Bonet,Kevin P. White,Adalgisa Caccone,Carlos López-Otín +35 more
TL;DR: The genomes of two long-lived giant tortoises, including Lonesome George, reveal candidate genes and pathways associated with their development, gigantism and longevity and expands the understanding of the genomic determinants of ageing.
Journal ArticleDOI
An Analysis of MIF Structural Features that Control Functional Activation of CD74.
Georgios Pantouris,Mansoor Ali Syed,Chengpeng Fan,Deepa Rajasekaran,Thomas Yoonsang Cho,Eric M. Rosenberg,Richard Bucala,Vineet Bhandari,Elias Lolis +8 more
TL;DR: Using a novel approach, this work elucidates the mechanistic details that control activation of CD74 by MIF surface residues and identifies structural parameters of inhibitors that reduce CD74 biological activation.
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Iterative model-building, structure refinement, and density modification with the PHENIX AutoBuild Wizard
TL;DR: The highly automated PHENIX AutoBuild wizard is described, which can be applied equally well to phases derived from isomorphous/anomalous and molecular-replacement methods.
Journal ArticleDOI
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Thierry Calandra,Jürgen Bernhagen,Christine N. Metz,Lori Spiegel,Michael Bacher,Thomas M. Donnelly,Anthony Cerami,Richard Bucala +7 more
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