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A 16-Residue Peptide Fragment of Macrophage Migration Inhibitory Factor, MIF-(50-65), Exhibits Redox Activity and Has MIF-like Biological Functions

Author: Nguyen, M.T. · Beck, J. · Lue, H. · Fünfzig, H. · Kleemann, R. · Koolwijk, P. · Kapurniotu, A. · Bernhagen, J.
Type:article
Date:2003
Institution: Gaubius Instituut TNO
Source:Journal of Biological Chemistry, 36, 278, 33654-33671
Identifier: 237277
doi: doi:10.1074/jbc.M301735200
Keywords: Biology · Biomedical Research · Cells · Conformations · Enzymes · Molecular biology · Redox reactions · Phosphorylation · Biochemistry · Animalia · 3T3 Cells · Amino Acid Motifs · Amino Acid Sequence · Animals · Biotinylation · Catalysis · Cell Cycle Proteins · Cell Division · Cells, Cultured · Chromatography, High Pressure Liquid · Circular Dichroism · Cyclin-Dependent Kinase Inhibitor p27 · Cysteine · Disulfides · Endocytosis · Endothelium, Vascular · Glutathione · Humans · Inflammation · Kinetics · Macrophage Migration-Inhibitory Factors · Mass Spectrometry · Mice · Mitogen-Activated Protein Kinase 1 · Mitogen-Activated Protein Kinase 3 · Mitogen-Activated Protein Kinases · Molecular Sequence Data · Oxidation-Reduction · Oxygen · Peptides · Phosphorylation · Protein Binding · Protein Conformation · Serine · Signal Transduction · Sulfhydryl Compounds · Time Factors · Tumor Suppressor Proteins

Abstract

Macrophage migration inhibitory factor (MIF) is a cytokine that participates in the host inflammatory response. A Cys-Xaa-Xaa-Cys (CXXC)-based thiol-protein oxidoreductase activity of MIF is associated with certain biological functions. Peptides spanning the CXXC region of thiol-protein oxidoreductases retain some biochemical properties of the full-length protein. We report on the characterization of CXXC-spanning MIF-(50-65) and its serine variant, C57S/C60S-MIF-(50-65). Following disulfide-mediated cyclization, MIF-(50-65) adapted a β-turn conformation comparable with that of β-turn-containing cyclo-57,60-[Asp57,Dap 60]MIF-(50-65). MIF-(50-65) had a redox potential E′ 0 of -0.258 V and formed mixed disulfides with glutathione and cysteine. MIF-(50-65) but not C57S/C60S-MIF-(50-65) had oxidoreductase activity in vitro. Intriguingly, MIF-(50-65) exhibited MIF-like cellular activities. The peptide but not its variant had glucocorticoid overriding and proliferation-enhancing activity and stimulated ERK1/2 phosphorylation. MIF-(50-65) and its variant bound to the MIF-binding protein JAB1 and enhanced cellular levels of p27Kip1. As the peptide and its variant were endocytosed at similar efficiency, sequence 50-65 appears sufficient for the JAB1-related effects of MIF, whereas other activities require CXXC. Cyclo-57,60-[Asp57,Dap60]MIF-(50-65) activated ERK1/2, indicating that CXXC-dependent disulfide and β-turn formation is associated with an activity-inducing conformation. We conclude that CXXC and sequence 50-65 are critical for the activities of MIF. MIF-(50-65) is a surprisingly short sequence with MIF-like functions that could be an excellent molecular template for MIF therapeutics.