6Q3S

Engineered Human HLA_A2 MHC Class I molecule in complex with TCR and SV9 peptide

Summary for 6Q3S

• Entry DOI: 10.2210/pdb6q3s/pdb
• Related: 2BNQ
• Descriptor: HLA class I histocompatibility antigen, A-2 alpha chain, Beta-2-microglobulin, SER-LEU-LEU-MET-TRP-ILE-THR-GLN-VAL, ... (8 entities in total)
• Functional Keywords: mhc molecule, engineered disulphide bond, immune system, tcr
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 5
• Total formula weight: 94622.03

Authors

Meijers, R.,Anjanappa, R.,Springer, S.,Garcia-Alai, M. (deposition date: 2018-12-04, release date: 2019-07-24, Last modification date: 2021-02-03)

Primary citation

Moritz, A.,Anjanappa, R.,Wagner, C.,Bunk, S.,Hofmann, M.,Pszolla, G.,Saikia, A.,Garcia-Alai, M.,Meijers, R.,Rammensee, H.G.,Springer, S.,Maurer, D.
High-throughput peptide-MHC complex generation and kinetic screenings of TCRs with peptide-receptive HLA-A*02:01 molecules.
Sci Immunol, 4:-, 2019

PubMed Abstract: Major histocompatibility complex (MHC) class I molecules present short peptide ligands on the cell surface for interrogation by cytotoxic CD8 T cells. MHC class I complexes presenting tumor-associated peptides such as neoantigens represent key targets of cancer immunotherapy approaches currently in development, making them important for efficacy and safety screenings. Without peptide ligand, MHC class I complexes are unstable and decay quickly, making the production of soluble monomers for analytical purposes labor intensive. We have developed a disulfide-stabilized HLA-A*02:01 molecule that is stable without peptide but can form peptide-MHC complexes (pMHCs) with ligands of choice in a one-step loading procedure. We illustrate the similarity between the engineered mutant and the wild-type molecule with respect to affinity of wild-type or affinity-matured T cell receptors (TCRs) and present a crystal structure corroborating the binding kinetics measurements. In addition, we demonstrate a high-throughput binding kinetics measurement platform to analyze the binding characteristics of bispecific TCR (bsTCR) molecules against diverse pMHC libraries produced with the disulfide-stabilized HLA-A*02:01 molecule. We show that bsTCR affinities for pMHCs are indicative of in vitro function and generate a bsTCR binding motif to identify potential off-target interactions in the human proteome. These findings showcase the potential of the platform and the engineered HLA-A*02:01 molecule in the emerging field of pMHC-targeting biologics.

PubMed: 31324691
DOI: 10.1126/sciimmunol.aav0860

Experimental method

X-RAY DIFFRACTION (2.5 Å)