4EVG

Crystal Structure of MIF L46A mutant

4EVG の概要

• エントリーDOI: 10.2210/pdb4evg/pdb
• 関連するPDBエントリー: 4ETG 4EUI
• 分子名称: Macrophage migration inhibitory factor, SULFATE ION (3 entities in total)
• 機能のキーワード: keto-enol tautomerase, isomerase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Secreted: P14174
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 37323.18

構造登録者

Ashrafi, A.,Pojer, F.,Lashuel, H. (登録日: 2012-04-26, 公開日: 2012-10-03, 最終更新日: 2024-02-28)

主引用文献

El-Turk, F.,Fauvet, B.,Ashrafi, A.,Ouertatani-Sakouhi, H.,Cho, M.K.,Neri, M.,Cascella, M.,Rothlisberger, U.,Pojer, F.,Zweckstetter, M.,Lashuel, H.
Characterization of molecular determinants of the conformational stability of macrophage migration inhibitory factor: leucine 46 hydrophobic pocket.
Plos One, 7:e45024-e45024, 2012

PubMed 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.

PubMed: 23028743
DOI: 10.1371/journal.pone.0045024

実験手法

X-RAY DIFFRACTION (1.7 Å)