7UFJ

Structure of human MR1-ethylvanillin in complex with human MAIT A-F7 TCR

7UFJ の概要

• エントリーDOI: 10.2210/pdb7ufj/pdb
• 分子名称: Major histocompatibility complex class I-related gene protein, Beta-2-microglobulin, MAIT T-cell receptor alpha chain, ... (9 entities in total)
• 機能のキーワード: immune system, antigen presentation, t-cell receptor, mr1
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 189571.33

構造登録者

Wang, C.J.,Rossjohn, J.,Le Nours, J. (登録日: 2022-03-22, 公開日: 2022-12-14, 最終更新日: 2024-11-13)

主引用文献

Wang, C.J.H.,Awad, W.,Liu, L.,Mak, J.Y.W.,Veerapen, N.,Illing, P.T.,Purcell, A.W.,Eckle, S.B.G.,McCluskey, J.,Besra, G.S.,Fairlie, D.P.,Rossjohn, J.,Le Nours, J.
Quantitative affinity measurement of small molecule ligand binding to Major Histocompatibility Complex class-I related protein 1 MR1.
J.Biol.Chem., 298:102714-102714, 2022

PubMed Abstract: The Major Histocompatibility Complex class I-related protein 1 (MR1) presents small molecule metabolites, drugs, and drug-like molecules that are recognized by MR1-reactive T cells. While we have an understanding of how antigens bind to MR1 and upregulate MR1 cell surface expression, a quantitative, cell-free, assessment of MR1 ligand-binding affinity was lacking. Here, we developed a fluorescence polarization-based assay in which fluorescent MR1 ligand was loaded into MR1 protein in vitro and competitively displaced by candidate ligands over a range of concentrations. Using this assay, ligand affinity for MR1 could be differentiated as strong (IC < 1 μM), moderate (1 μM < IC < 100 μM), and weak (IC > 100 μM). We demonstrated a clear correlation between ligand-binding affinity for MR1, the presence of a covalent bond between MR1 and ligand, and the number of salt bridge and hydrogen bonds formed between MR1 and ligand. Using this newly developed fluorescence polarization-based assay to screen for candidate ligands, we identified the dietary molecules vanillin and ethylvanillin as weak bona fide MR1 ligands. Both upregulated MR1 on the surface of C1R.MR1 cells and the crystal structure of a MAIT cell T cell receptor-MR1-ethylvanillin complex revealed that ethylvanillin formed a Schiff base with K43 of MR1 and was buried within the A'-pocket. Collectively, we developed and validated a method to quantitate the binding affinities of ligands for MR1 that will enable an efficient and rapid screening of candidate MR1 ligands.

PubMed: 36403855
DOI: 10.1016/j.jbc.2022.102714

実験手法

X-RAY DIFFRACTION (2.5 Å)