Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor

Leucine 46 Hydrophobic Pocket

Farah El-Turk, Bruno Fauvet, Amer Ashrafi, Hajer Ouertatani-Sakouhi, Min Kyu Cho, Marilisa Neri, Michele Cascella, Ursula Rothlisberger, Florence Pojer, Markus Zweckstetter, Hilal Lashuel

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Macrophage Migration Inhibitory Factor (MIF) is a key mediator of inflammatory responses and innate immunity and has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. The oligomerization of MIF, more specifically trimer formation, is essential for its keto-enol tautomerase activity and probably mediates several of its interactions and biological activities, including its binding to its receptor CD74 and activation of certain signaling pathways. Therefore, understanding the molecular factors governing the oligomerization of MIF and the role of quaternary structure in modulating its structural stability and multifunctional properties is crucial for understanding the function of MIF in health and disease. Herein, we describe highly conserved intersubunit interactions involving the hydrophobic packing of the side chain of Leu46 onto the β-strand β3 of one monomer within a hydrophobic pocket from the adjacent monomer constituted by residues Arg11, Val14, Phe18, Leu19, Val39, His40, Val41, Val42, and Pro43. To elucidate the structural significance of these intersubunit interactions and their relative contribution to MIF's trimerization, structural stability and catalytic activity, we generated three point mutations where Leu46 was replaced by glycine (L46G), alanine (L46A) and phenylalanine (L46F), and their structural properties, stability, oligomerization state, and catalytic activity were characterized using a battery of biophysical methods and X-ray crystallography. Our findings provide new insights into the role of the Leu46 hydrophobic pocket in stabilizing the conformational state of MIF in solution. Disrupting the Leu46 hydrophobic interaction perturbs the secondary and tertiary structure of the protein but has no effect on its oligomerization state.

Original languageEnglish
Article numbere45024
JournalPLoS One
Volume7
Issue number9
DOIs
Publication statusPublished - 21 Sep 2012
Externally publishedYes

Fingerprint

Macrophage Migration-Inhibitory Factors
Oligomerization
Hydrophobic and Hydrophilic Interactions
Leucine
leucine
Secondary Protein Structure
hydrophobic bonding
X Ray Crystallography
Phenylalanine
Tertiary Protein Structure
catalytic activity
Point Mutation
Innate Immunity
Alanine
Glycine
Autoimmune Diseases
Catalyst activity
Monomers
protein tertiary structure
autoimmune diseases

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor : Leucine 46 Hydrophobic Pocket. / El-Turk, Farah; Fauvet, Bruno; Ashrafi, Amer; Ouertatani-Sakouhi, Hajer; Cho, Min Kyu; Neri, Marilisa; Cascella, Michele; Rothlisberger, Ursula; Pojer, Florence; Zweckstetter, Markus; Lashuel, Hilal.

In: PLoS One, Vol. 7, No. 9, e45024, 21.09.2012.

Research output: Contribution to journalArticle

El-Turk, F, Fauvet, B, Ashrafi, A, Ouertatani-Sakouhi, H, Cho, MK, Neri, M, Cascella, M, Rothlisberger, U, Pojer, F, Zweckstetter, M & Lashuel, H 2012, 'Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor: Leucine 46 Hydrophobic Pocket', PLoS One, vol. 7, no. 9, e45024. https://doi.org/10.1371/journal.pone.0045024
El-Turk, Farah ; Fauvet, Bruno ; Ashrafi, Amer ; Ouertatani-Sakouhi, Hajer ; Cho, Min Kyu ; Neri, Marilisa ; Cascella, Michele ; Rothlisberger, Ursula ; Pojer, Florence ; Zweckstetter, Markus ; Lashuel, Hilal. / Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor : Leucine 46 Hydrophobic Pocket. In: PLoS One. 2012 ; Vol. 7, No. 9.
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AU - Ashrafi, Amer

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AU - Cho, Min Kyu

AU - Neri, Marilisa

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