Title
A 16-Residue Peptide Fragment of Macrophage Migration Inhibitory Factor, MIF-(50-65), Exhibits Redox Activity and Has MIF-like Biological Functions
Author
Gaubius Instituut TNO
Nguyen, M.T.
Beck, J.
Lue, H.
Fünfzig, H.
Kleemann, R.
Koolwijk, P.
Kapurniotu, A.
Bernhagen, J.
Publication year
2003
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 beta-turn conformation comparable with that of beta-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 beta-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.
Subject
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
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DOI
https://doi.org/10.1074/jbc.m301735200
TNO identifier
237277
ISSN
0021-9258
Source
Journal of Biological Chemistry, 278 (278), 33654-33671
Document type
article