A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor

Hajer Ouertatani-Sakouhi, Farah El-Turk, Bruno Fauvet, Thierry Roger, Didier Le Roy, Damla Pinar Karpinar, Lin Leng, Richard Bucala, Markus Zweckstetter, Thierry Calandra, Hilal A. Lashuel

Research output: Contribution to journalArticle

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Abstract

Macrophage migration inhibitory factor (MIF) is a homotrimeric multifunctional proinflammatory cytokine that has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. Current therapeutic strategies for targeting MIF focus on developing inhibitors of its tautomerase activity or modulating its biological activities using anti-MIF neutralizing antibodies. Herein we report a new class of isothiocyanate (ITC)-based irreversible inhibitors of MIF. Modification by benzyl isothiocyanate (BITC) and related analogues occurred at the N-terminal catalytic proline residue without any effect on the oligomerization state of MIF. Different alkyl and arylalkyl ITCs modified MIF with nearly the same efficiency as BITC. To elucidate the mechanism of action, we performed detailed biochemical, biophysical, and structural studies to determine the effect of BITC and its analogues on the conformational state, quaternary structure, catalytic activity, receptor binding, and biological activity of MIF. Light scattering, analytical ultracentrifugation, and NMR studies on unmodified and ITC-modified MIF demonstrated that modification of Pro1 alters the tertiary, but not the secondary or quaternary, structure of the trimer without affecting its thermodynamic stability. BITC induced drastic effects on the tertiary structure of MIF, in particular residues that cluster around Pro1 and constitute the tautomerase active site. These changes in tertiary structure and the loss of catalytic activity translated into a reduction in MIF receptor binding activity, MIF-mediated glucocorticoid overriding, and MIF-induced Akt phosphorylation. Together, these findings highlight the role of tertiary structure in modulating the biochemical and biological activities of MIF and present new opportunities for modulating MIF biological activities in vivo.

Original languageEnglish
Pages (from-to)9858-9870
Number of pages13
JournalBiochemistry
Volume48
Issue number41
DOIs
Publication statusPublished - 20 Oct 2009
Externally publishedYes

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Macrophage Migration-Inhibitory Factors
Bioactivity
Catalyst activity
Oligomerization
Phosphorylation
Ultracentrifugation
Biological Factors
Neutralizing Antibodies
Thermodynamics
Proline
Light scattering
Glucocorticoids
Autoimmune Diseases
Catalytic Domain
Thermodynamic stability
Nuclear magnetic resonance
Cytokines
Light
benzyl isothiocyanate
isothiocyanic acid

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ouertatani-Sakouhi, H., El-Turk, F., Fauvet, B., Roger, T., Roy, D. L., Karpinar, D. P., ... Lashuel, H. A. (2009). A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor. Biochemistry, 48(41), 9858-9870. https://doi.org/10.1021/bi900957e

A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor. / Ouertatani-Sakouhi, Hajer; El-Turk, Farah; Fauvet, Bruno; Roger, Thierry; Roy, Didier Le; Karpinar, Damla Pinar; Leng, Lin; Bucala, Richard; Zweckstetter, Markus; Calandra, Thierry; Lashuel, Hilal A.

In: Biochemistry, Vol. 48, No. 41, 20.10.2009, p. 9858-9870.

Research output: Contribution to journalArticle

Ouertatani-Sakouhi, H, El-Turk, F, Fauvet, B, Roger, T, Roy, DL, Karpinar, DP, Leng, L, Bucala, R, Zweckstetter, M, Calandra, T & Lashuel, HA 2009, 'A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor', Biochemistry, vol. 48, no. 41, pp. 9858-9870. https://doi.org/10.1021/bi900957e
Ouertatani-Sakouhi H, El-Turk F, Fauvet B, Roger T, Roy DL, Karpinar DP et al. A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor. Biochemistry. 2009 Oct 20;48(41):9858-9870. https://doi.org/10.1021/bi900957e
Ouertatani-Sakouhi, Hajer ; El-Turk, Farah ; Fauvet, Bruno ; Roger, Thierry ; Roy, Didier Le ; Karpinar, Damla Pinar ; Leng, Lin ; Bucala, Richard ; Zweckstetter, Markus ; Calandra, Thierry ; Lashuel, Hilal A. / A new class of isothiocyanate-based irreversible inhibitors of macrophage migration inhibitory factor. In: Biochemistry. 2009 ; Vol. 48, No. 41. pp. 9858-9870.
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AU - Karpinar, Damla Pinar

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AB - Macrophage migration inhibitory factor (MIF) is a homotrimeric multifunctional proinflammatory cytokine that has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. Current therapeutic strategies for targeting MIF focus on developing inhibitors of its tautomerase activity or modulating its biological activities using anti-MIF neutralizing antibodies. Herein we report a new class of isothiocyanate (ITC)-based irreversible inhibitors of MIF. Modification by benzyl isothiocyanate (BITC) and related analogues occurred at the N-terminal catalytic proline residue without any effect on the oligomerization state of MIF. Different alkyl and arylalkyl ITCs modified MIF with nearly the same efficiency as BITC. To elucidate the mechanism of action, we performed detailed biochemical, biophysical, and structural studies to determine the effect of BITC and its analogues on the conformational state, quaternary structure, catalytic activity, receptor binding, and biological activity of MIF. Light scattering, analytical ultracentrifugation, and NMR studies on unmodified and ITC-modified MIF demonstrated that modification of Pro1 alters the tertiary, but not the secondary or quaternary, structure of the trimer without affecting its thermodynamic stability. BITC induced drastic effects on the tertiary structure of MIF, in particular residues that cluster around Pro1 and constitute the tautomerase active site. These changes in tertiary structure and the loss of catalytic activity translated into a reduction in MIF receptor binding activity, MIF-mediated glucocorticoid overriding, and MIF-induced Akt phosphorylation. Together, these findings highlight the role of tertiary structure in modulating the biochemical and biological activities of MIF and present new opportunities for modulating MIF biological activities in vivo.

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