Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode
Abstract
Natural T cell receptors (TCRs) generally bind to their cognate pMHC molecules with weak affinity and fast kinetics, limiting their use as therapeutic agents. Using phage display, we have engineered a high affinity version of the A6 wild-type TCR (A6wt), specific for the human leukocyte antigen (HLA-A*0201) complexed with human T cell lymphotropic virus type 111–19 peptide (A2-Tax). Mutations in just 4 residues in the CDR3β loop region of the A6wt TCR were selected that improved binding to A2-Tax by nearly 1000-fold. Biophysical measurements of this mutant TCR (A6c134) demonstrated that the enhanced binding was derived through favorable enthalpy and a slower off-rate. The structure of the free A6c134 TCR and the A6c134/A2-Tax complex revealed a native binding mode, similar to the A6wt/A2-Tax complex. However, concordant with the more favorable binding enthalpy, the A6c134TCR made increased contacts with the Tax peptide compared with the A6wt/A2- Tax complex, demonstrating a peptide-focused mechanism for the enhanced affinity that directly involved the mutated residues in the A6c134TCR CDR3β loop. This peptide-focused enhanced TCR binding may represent an important approach for developing antigen specific high affinity TCR reagents for use in T cell based therapies.
- Authors:
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
- OSTI Identifier:
- 1628044
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Frontiers in Immunology
- Additional Journal Information:
- Journal Volume: 4; Journal ID: ISSN 1664-3224
- Publisher:
- Frontiers Research Foundation
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Immunology; human T leukocyte virus type 1; crystal structure; peptide-major histocompatibility complex; surface plasmon resonance; T cell; T cell receptor; A6 TCR; high affinity TCR
Citation Formats
Cole, David. Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode. United States: N. p., 2013.
Web. doi:10.3389/fimmu.2013.00168.
Cole, David. Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode. United States. https://doi.org/10.3389/fimmu.2013.00168
Cole, David. Tue .
"Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode". United States. https://doi.org/10.3389/fimmu.2013.00168. https://www.osti.gov/servlets/purl/1628044.
@article{osti_1628044,
title = {Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode},
author = {Cole, David},
abstractNote = {Natural T cell receptors (TCRs) generally bind to their cognate pMHC molecules with weak affinity and fast kinetics, limiting their use as therapeutic agents. Using phage display, we have engineered a high affinity version of the A6 wild-type TCR (A6wt), specific for the human leukocyte antigen (HLA-A*0201) complexed with human T cell lymphotropic virus type 111–19 peptide (A2-Tax). Mutations in just 4 residues in the CDR3β loop region of the A6wt TCR were selected that improved binding to A2-Tax by nearly 1000-fold. Biophysical measurements of this mutant TCR (A6c134) demonstrated that the enhanced binding was derived through favorable enthalpy and a slower off-rate. The structure of the free A6c134 TCR and the A6c134/A2-Tax complex revealed a native binding mode, similar to the A6wt/A2-Tax complex. However, concordant with the more favorable binding enthalpy, the A6c134TCR made increased contacts with the Tax peptide compared with the A6wt/A2- Tax complex, demonstrating a peptide-focused mechanism for the enhanced affinity that directly involved the mutated residues in the A6c134TCR CDR3β loop. This peptide-focused enhanced TCR binding may represent an important approach for developing antigen specific high affinity TCR reagents for use in T cell based therapies.},
doi = {10.3389/fimmu.2013.00168},
journal = {Frontiers in Immunology},
number = ,
volume = 4,
place = {United States},
year = {Tue Jan 01 00:00:00 EST 2013},
month = {Tue Jan 01 00:00:00 EST 2013}
}
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