Quantitative affinity measurement of small molecule ligand binding to major histocompatibility complex class-I–related protein 1 MR1

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 (IC50 < 1 μM), moderate (1 μM < IC50 < 100 μM), and weak (IC50 > 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 Aʹ-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. 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 (IC50 < 1 μM), moderate (1 μM < IC50 < 100 μM), and weak (IC50 > 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 Aʹ-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. In vertebrates, protective immunity against pathogens relies heavily on a specialized class of molecules, encoded by the Major Histocompatibility Complex (MHC), which present pathogen-derived peptides to T cells for recognition and elimination. In jawed vertebrates, there exist three T cell lineages, namely, αβ, γδ, γμ T cells, which are defined by the cell surface expression of T cell receptor (TCR) heterodimers composed of α- and β- or γ- and δ- or γ- and μ-polypeptide chains, respectively (1Morrissey K.A. Wegrecki M. Praveena T. Hansen V.,L. Bu L. Sivaraman K.,K. et al.The molecular assembly of the marsupial γμ T cell receptor defines a third T cell lineage.Science. 2021; 371: 1383-1388Crossref PubMed Scopus (10) Google Scholar, 2Chien Y. Becker D.M. Lindsten T. Okamura M. Cohen D.I. Davis M.M. A third type of murine T-cell receptor gene.Nature. 1984; 312: 31-35Crossref PubMed Scopus (367) Google Scholar, 3Yanagi Y. Yoshikai Y. Leggett K. Clark S.P. Aleksander I. Mak T.W. A human T cell-specific cDNA clone encodes a protein having extensive homology to immunoglobulin chains.Nature. 1984; 308: 145-149Crossref PubMed Scopus (888) Google Scholar, 4Brenner M.B. McLean J. Dialynas D.P. Strominger J.L. Smith J.A. Owen F.L. et al.Identification of a putative second T-cell receptor.Nature. 1986; 322: 145-149Crossref PubMed Scopus (724) Google Scholar). The fundamental principles underscoring the mechanism of recognition of the MHC-peptide complex by αβ TCRs have been extensively studied over the past 25 years (5Rossjohn J. Gras S. Miles J.J. Turner S.J. Godfrey D.I. McCluskey J. T cell antigen receptor recognition of antigen-presenting molecules.Annu. Rev. Immunol. 2015; 33: 169-200Crossref PubMed Scopus (461) Google Scholar). In addition to classical MHC-I and MHC-II molecules, MHC-I–like molecules exist that include the MHC-I related protein 1 (MR1) and the CD1 family of glycoproteins that present small molecules and lipid-derived antigens (Ags) to T cells, respectively (5Rossjohn J. Gras S. Miles J.J. Turner S.J. Godfrey D.I. McCluskey J. T cell antigen receptor recognition of antigen-presenting molecules.Annu. Rev. Immunol. 2015; 33: 169-200Crossref PubMed Scopus (461) Google Scholar). Such MHC-I–like molecules can be recognized by specialized immune cell subsets that include αβ and γδ T cells (5Rossjohn J. Gras S. Miles J.J. Turner S.J. Godfrey D.I. McCluskey J. T cell antigen receptor recognition of antigen-presenting molecules.Annu. Rev. Immunol. 2015; 33: 169-200Crossref PubMed Scopus (461) Google Scholar, 6Rice M.T. von Borstel A. Chevour P. Awad W. Howson L.J. Littler D.R. et al.Recognition of the antigen-presenting molecule MR1 by a Vδ3(+) γδ T cell receptor.Proc. Natl. Acad. Sci. U. S. A. 2021; 118e2110288118Crossref Scopus (12) Google Scholar, 7Le Nours J. Gherardin N.A. Ramarathinam S.H. Awad W. Wiede F. Gully B.S. et al.A class of γδ T cell receptors recognize the underside of the antigen-presenting molecule MR1.Science. 2019; 366: 1522-1527Crossref PubMed Scopus (69) Google Scholar, 8Willcox B.E. Willcox C.R. Γδ TCR ligands: the quest to solve a 500-million-year-old mystery.Nat. Immunol. 2019; 20: 121-128Crossref PubMed Scopus (82) Google Scholar). The monomorphic MR1 molecule is ubiquitously in human cells and with the classical MHC-I molecules, to a class of namely, and L. Nours J. L. et presents to MAIT PubMed Scopus Google Scholar, L. J. et by PubMed Scopus Google that are recognized by of T cells, a T cells Godfrey D.I. L. et al.Recognition of and by T 2015; PubMed Scopus Google and αβ and γδ T cells M.T. von Borstel A. Chevour P. Awad W. Howson L.J. Littler D.R. et al.Recognition of the antigen-presenting molecule MR1 by a Vδ3(+) γδ T cell receptor.Proc. Natl. Acad. Sci. U. S. A. 2021; 118e2110288118Crossref Scopus (12) Google Scholar, 7Le Nours J. Gherardin N.A. Ramarathinam S.H. Awad W. Wiede F. Gully B.S. et al.A class of γδ T cell receptors recognize the underside of the antigen-presenting molecule MR1.Science. 2019; 366: 1522-1527Crossref PubMed Scopus (69) Google Scholar, N.A. Nours J. et of T by the class molecule MR1 antigen PubMed Scopus Google Scholar, M. A. S. P. M. et human T cells recognize antigens by Scholar, M. A. M. et screening T cell the monomorphic class protein Immunol. PubMed Scopus Google Scholar, Gherardin N.A. et T cells in and 2019; PubMed Scopus Google Scholar). The structure and of the MR1 are to bind small molecules, the ligands are by a of and that the L. Nours J. L. et presents to MAIT PubMed Scopus Google Scholar). the of MR1 ligands or a covalent is formed between K43 of MR1 and the or L. Nours J. L. et presents to MAIT PubMed Scopus Google Scholar). The of the is to an of of and ligands a Schiff base with within the the of the ligands to with and MR1 hydrogen of the to the of the for W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, M. M. et the T cell receptor Immunol. PubMed Scopus Google the of molecules that can be by MR1 and MAIT cells L. D.P. of T 2021; PubMed Scopus Google Scholar). have been in understanding the of MR1 Mak Awad W. M. S. et MR1 molecules for efficient of Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, A. L. et al.The for the of antigens by the antigen-presenting molecule Immunol. PubMed Scopus Google Scholar, A. et MR1 a in of ligands to MAIT PubMed Scopus Google the molecular antigen by MR1, as as MAIT TCR and of ligands W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar). into of ligand affinity for MR1 have been been to the binding affinity of peptides for MHC-I and MHC-II L. L.J. of binding to class molecules by fluorescence Immunol. PubMed Scopus Google Scholar, S. S. S. of fluorescence to in Immunol. PubMed Scopus Google Scholar). Here, we this by a fluorescent ligand Mak Awad W. M. S. et MR1 molecules for efficient of Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, J. Ramarathinam S.H. et al.The between and in binding in PubMed Scopus Google to a assay to the binding affinity of ligands to MR1 and the molecular that affinity The assay the of ligands and as MR1 ligands. The crystal structure of a MAIT TCR in complex with how ethylvanillin was within the of MR1, to bind assay a and to the affinity of candidate ligands for MR1 and be a screening While we have an understanding of how bind to MR1 and upregulate MR1 cell surface expression, the affinity of ligands binding to MR1 are to be The affinity of ligands was to and in W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, L. et al.A molecular the T cell receptor of T PubMed Scopus Google Scholar, Y. et a mechanism to MR1 PubMed Scopus Google Scholar). is a of ligand binding and is to ligands that have the to be with MR1 in on was to L. L.J. of binding to class molecules by fluorescence Immunol. PubMed Scopus Google Scholar, J. Ramarathinam S.H. et al.The between and in binding in PubMed Scopus Google we to an assay for the of the affinity of MR1 ligands for MR1 in a In assay, we a fluorescent of the MAIT cell Mak Awad W. M. S. et MR1 molecules for efficient of Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). fluorescent that with MR1 been that competitively of MAIT cells and that to MR1 to the binding of Mak Awad W. M. S. et MR1 molecules for efficient of Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). into MR1 in we an MR1 molecule and the in vitro of MR1 with the and W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google of the Schiff base between and K43 of MR1 A. L. et al.The for the of antigens by the antigen-presenting molecule Immunol. PubMed Scopus Google the of the been to the structure of and drug-like molecules binding to MR1 W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). The was and the molecule was by The molecule the for The and of MR1 and and was a MR1 can be in and was as an and efficient for affinity the for a binding assay of and MR1 was to the of MR1 and to a as as the to of the is by that will be and as is in as the molecule in a The is is to MR1, to the of to the MR1 protein D.M. J.A. in and Rev. PubMed Scopus Google Scholar). The of a MR1 molecule and the of a fluorescent ligand for the of an assay to the affinity of a range of MR1 ligands Here, a of 25 and was was to as 25 and 25 in which MR1 over of to a of we that a of a of 100 and of and and an of 25 to the to within and the MR1 molecule to A was to the to the of MR1, an the MHC-I molecule S. K. A. et for the of a TCR against a human Immunol. PubMed Scopus Google was as a and was to bind to we the assay could between MR1 and We as the ligand of that was to bind to MR1 L. et al.A molecular the T cell receptor of T PubMed Scopus Google and the S. K. A. et for the of a TCR against a human Immunol. PubMed Scopus Google as a that is to be displaced the MR1 by ligand by In this identified the for an assay for MR1 ligand and demonstrated for MR1 ligands. Using the assay we the affinity of a range of of MR1 ligands 1 and we ligands that the ligands: L. Nours J. L. et presents to MAIT PubMed Scopus Google and L. et al.A molecular the T cell receptor of T PubMed Scopus Google the ligands: the of and L. J. et by PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, W. B.S. et Schiff base of that T Scopus Google the ligands: and L. Nours J. L. et presents to MAIT PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google as as and the ligands and W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google ligand affinities of ligands binding to moderate and weak binding three in are with in for ligands are in MR1, protein of μM μM μM μM μM μM μM in a the and ligands that formed a Schiff base with MR1 the affinity for MR1 with IC50 < 1 μM the and to be the of the covalent bond for affinity MR1 this the MAIT cell by the affinity (IC50 by and with an to in affinity to the the have in the the in IC50 the of the for MAIT TCR W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google for MR1 an in affinity was between and is with the to the which in the of to with MR1 as W. Mak L. D.P. and of and of a that human T J. 2019; Scopus Google Scholar). strong of MR1 cell surface expression, and affinity (IC50 and for MR1 that was to the and are and bind to and a have a to a covalent bond with K43 of MR1 as as a that hydrogen bonds and to for the of on the of the (1 μM < IC50 < 100 to (IC50 are the base ligands that include the and and and ligands to surface MR1 expression and M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). Here, the ligands a or and a the of Schiff base with K43 to bind to MR1 with IC50 between to 100 the ligand a Schiff base with MR1, a binding affinity (IC50 to the number of hydrogen bonds and W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). Collectively, the that the of a ligand to a covalent bond with MR1 affinity for between MR1 and ligand can binding affinity for ligands. in screening identified vanillin and ethylvanillin as molecules with hydrogen and an that could enable binding to MR1 W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). this we the assay to binding affinities for and ethylvanillin to have affinities for MR1 with to μM as weak ligands to the of surface MR1 on the human cell MR1 we MR1 a with ethylvanillin and vanillin Here, we that ethylvanillin of MR1, vanillin a in MR1 that small molecules can be within a we for MR1 a Here, we demonstrated that surface MR1 was upregulated a with ethylvanillin and vanillin While the affinity for MR1, vanillin upregulated MR1 a that of ethylvanillin that the of the in to the cells, and to be by MR1 on the cell into the molecular of ethylvanillin by MR1, a was to ethylvanillin in vitro into W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). Using of the MAIT complex we a that to was an to the ethylvanillin the presence of ethylvanillin within the MAIT we the crystal structure of the MAIT complex to and The the and for the ethylvanillin was and The by the MAIT TCR to recognize the complex was to MAIT W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, L. et al.A molecular the T cell receptor of T PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google Here, the TCR a and the formed by the and The and the the with a of and to the buried surface The of the molecular the with in MAIT The ethylvanillin was buried within the of the and was for MAIT TCR formed a Schiff base with the K43 of to ligands, ethylvanillin was within an formed by and and While the of ethylvanillin with and the of the of ethylvanillin formed and hydrogen bonds with and respectively binding of there was of the the of MR1 to the crystal of W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, L. et al.A molecular the T cell receptor of T PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google Scholar). ethylvanillin an that was Schiff ligands as and that ethylvanillin is the of MR1 in to ligands. the of the to a range of ligands the of the binding we the that the affinity for the ligands. We the crystal of the MAIT TCR in complex with MR1 of ligands and W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, L. et al.A molecular the T cell receptor of T PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google to the number of molecular that hydrogen bonds salt and and between MR1 and the ligand. against the binding affinity of ligand to for the Schiff and ligands In three that the for Schiff and ligands with and the of Schiff base to ligand a correlation was between the number of and the affinity of a ligand with correlation of of base and base The Schiff and ligands a with and ligand affinity and correlation with the and the of the ligand and the ligands a with and and with a moderate correlation with and that the of ligand affinities the assay with and that ligand affinity is on the number and of between ligand and The of MR1 is to a range of of ligands recognized by MAIT cells and subsets of MR1-reactive T cells M.T. von Borstel A. Chevour P. Awad W. Howson L.J. Littler D.R. et al.Recognition of the antigen-presenting molecule MR1 by a Vδ3(+) γδ T cell receptor.Proc. Natl. Acad. Sci. U. S. A. 2021; 118e2110288118Crossref Scopus (12) Google Scholar, 7Le Nours J. Gherardin N.A. Ramarathinam S.H. Awad W. Wiede F. Gully B.S. et al.A class of γδ T cell receptors recognize the underside of the antigen-presenting molecule MR1.Science. 2019; 366: 1522-1527Crossref PubMed Scopus (69) Google Scholar, N.A. Nours J. et of T by the class molecule MR1 antigen PubMed Scopus Google Scholar, M. A. S. P. M. et human T cells recognize antigens by Scholar, M. A. M. et screening T cell the monomorphic class protein Immunol. PubMed Scopus Google Scholar, Gherardin N.A. et T cells in and 2019; PubMed Scopus Google Scholar). there is method for ligand affinity for for molecules, MHC-I and MHC-II L. L.J. of binding to class molecules by fluorescence Immunol. PubMed Scopus Google have been for in L. L.J. of binding to class molecules by fluorescence Immunol. PubMed Scopus Google W. J. et structure of the complex defines for rapid PubMed Scopus Google and K.A. a mechanism that hydrogen bonds in class MHC-peptide Immunol. PubMed Scopus Google Scholar). with a of that molecules bind and the affinities with which the peptides bind to molecules can be J. Ramarathinam S.H. et al.The between and in binding in PubMed Scopus Google Scholar). Here, we have developed an assay for the binding affinities of putative ligands for MR1, and of MAIT cells. In have been to the crystal structure and which is and developed assay the and of the affinity of ligands for the MR1 an efficient and rapid screening method for ligands that can be for of MAIT cell or We demonstrated that the ligand-binding assay is for the of ligands with IC50 in the to assay the clear correlation between Schiff base the number of covalent and the in binding affinity of a ligand. The affinity of a for within the to the μM range W. and of a fluorescence assay for for PubMed Scopus Google Scholar). While peptides weak to moderate affinity for are and an immune in the of T cell the to the MHC-II with a weak affinity J. Ramarathinam S.H. et al.The between and in binding in PubMed Scopus Google and can an in MHC-II J.A. W. et by in PubMed Scopus Google Scholar). affinity MR1 ligand as (IC50 of have been to be to a MAIT TCR cell by a cell MR1 W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). the assay the of MR1 ligands as the dietary molecules vanillin and vanillin and are present in a in and L. method for the of and vanillin in by with PubMed Scopus Google Scholar). is and vanillin can be can be the of and is in as and S. M. L. L. et and of PubMed Scopus Google Scholar). the of ethylvanillin to bind within the of the the MAIT affinities for MR1, that a correlation between the affinity of a ligand for MR1 and to upregulate MR1 be the of a ligand to and the cell the in surface MR1 between ligands. that MAIT cells the and to of T cells M. I. et MAIT cells are T PubMed Scopus Google this as an screening for the of MR1 ligands the of into the to present within the that MR1-reactive T cells, of or dietary Collectively, this a and rapid method for ligands to of the of the ethylvanillin and by and by was by L. D.P. of T 2021; PubMed Scopus Google Mak Awad W. M. S. et MR1 molecules for efficient of Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. B.S. et Schiff base of that T Scopus Google W. Mak L. D.P. and of and of a that human T J. 2019; Scopus Google as W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, W. B.S. et Schiff base of that T Scopus Google the of W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google L. Nours J. L. et presents to MAIT PubMed Scopus Google W. Mak L. D.P. and of and of a that human T J. 2019; Scopus Google and W. B.S. et Schiff base of that T Scopus Google as The and ligands M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google by of was in and ligand was in and was in with the of MR1 for in the assay, was with a range of MR1 in assay was and 25 the The ligand-binding and as a the of for binding to The MR1 ligands to and with and MR1 in assay was and 25 the as a IC50 for binding affinity the ligand for of for binding to The of the crystal structure of the the and W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar, M. Awad W. et of MR1 cell surface Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, W. W. Mak et al.The molecular the and of MAIT cell Immunol. PubMed Scopus Google Scholar, L. et al.A molecular the T cell receptor of T PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google and for the The number of hydrogen salt and the of P. et of the and PubMed Scopus Google Scholar). A for for salt and for hydrogen bonds was was The was against the IC50 of ligand to a for the of MR1, and the and L. Nours J. L. et presents to MAIT PubMed Scopus Google Scholar, L. Nours J. T. et al.Recognition of by T PubMed Scopus Google Scholar, Gras S. and of in complex with 2021; PubMed Scopus Google into cells. The and as MR1 and the rapid and and as W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). of in of of and of and in a of of of of and for The was against and the complex was was to with and against for The was to and as W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google Scholar). cells MR1 L. M. et into recognition of the T cell PubMed Scopus Google with ligand for or and in 100 or with (1 to and The cells with or and with S. S. M. L. et for MR1 antigen to T PubMed Scopus Google Scholar). was with and the cells with or and in was with an and and and TCR and a of in a and μM of ethylvanillin was The complex in 100 to and The in with and in A was on the J. M. et A the and the PubMed Scopus Google to W. PubMed Scopus Google and the of P. et of the and PubMed Scopus Google Scholar). The complex structure was by molecular PubMed Scopus Google and the structure the ligand W. L. Mak et and drug-like molecules can the of T Immunol. PubMed Scopus Google as with P. K. and of PubMed Scopus Google and with J.J. M. et structure with PubMed Scopus Google the complex crystal structure to an of The of the structure was the for and of molecular with the molecular in ligands, of protein was with a and for protein was by for The was and the was by to the in The small molecules by an with a with a The was loaded a in for The was a an of in over The was in with the 1 25 The range was to 100 to with a of the for a of to > for of an ethylvanillin was by the and structure of the MAIT complex in the the J. L. P. S. J. and J. Y. W. M. are on MR1 and that have of with the of this We the the for with and the We for with the MR1 We and for in the of the MR1 binding The was on the the of J. M. et A the and the PubMed Scopus Google Scholar). We the and of in the and the and of and the of for J. W. J. W. L. J. Y. W. P. T. A. W. S. J. S. and P. F. L. J. Y. W. S. J. S. and P. F. J. J. and J. L. J. J. and J. L. J. W. is by an W. A. is by an L. J. Y. W. and P. F. by and A. W. P. is by an J. M. and J. are by S. is by an The is the of the and the of the of

Topics & Concepts