High-affinity, peptide-specific T cell receptors can be generated by mutations in CDR1, CDR2 or CDR3

Lukasz K. Chlewicki, Phillip D. Holler, Bridget C. Monti, Matthew R. Clutter, David M. Kranz

Research output: Contribution to journalArticle

Abstract

The third complementarity-determining regions (CDR3s) of antibodies and T cell receptors (TCRs) have been shown to play a major role in antigen binding and specificity. Consistent with this notion, we demonstrated previously that high-affinity, peptide-specific TCRs could be generated in vitro by mutations in the CDR3α region of the 2C TCR. In contrast, it has been argued that CDR1 and CDR2 are involved to a greater extent than CDR3s in the process of MHC restriction, due to their engagement of MHC helices. Based on this premise, we initiated the present study to explore whether higher affinity TCRs generated through mutations in these CDRs or other regions would lead to significant reductions in peptide specificity (i.e. the result of greater binding energy gained through interactions with major histocompatibility complex (MHC) helices). Yeast-display technology and flow sorting were used to select high-affinity TCRs from libraries of CDR mutants or random mutants. High-affinity TCRs with mutations in the first residue of the Vα, CDR1, CDR2, or CDR3 were isolated. Unexpectedly, every TCR mutant, including those in CDR1 and CDR2, retained remarkable peptide specificity. Molecular modeling of various mutants suggested that such exquisite specificity may be due to: (1) enhanced electrostatic interactions with key peptide or MHC residues; or (2) stabilization of CDRs in specific conformations. The results indicate that the TCR is positioned so that virtually every CDR can contribute to the antigen-specificity of a T cell. The conserved diagonal docking of TCRs could thus orient each CDR loop to sense the peptide directly or indirectly through peptide-induced effects on the MHC.

Original languageEnglish (US)
Pages (from-to)223-239
Number of pages17
JournalJournal of Molecular Biology
Volume346
Issue number1
DOIs
StatePublished - Feb 11 2005

Fingerprint

Peptide T
T-Cell Antigen Receptor
Mutation
Major Histocompatibility Complex
Peptides
T-Cell Antigen Receptor Specificity
Complementarity Determining Regions
Antigens
Static Electricity
Libraries
Yeasts
Technology

Keywords

  • T cell receptors
  • antigen recognition
  • protein engineering
  • yeast display

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

High-affinity, peptide-specific T cell receptors can be generated by mutations in CDR1, CDR2 or CDR3. / Chlewicki, Lukasz K.; Holler, Phillip D.; Monti, Bridget C.; Clutter, Matthew R.; Kranz, David M.

In: Journal of Molecular Biology, Vol. 346, No. 1, 11.02.2005, p. 223-239.

Research output: Contribution to journalArticle

Chlewicki, Lukasz K. ; Holler, Phillip D. ; Monti, Bridget C. ; Clutter, Matthew R. ; Kranz, David M. / High-affinity, peptide-specific T cell receptors can be generated by mutations in CDR1, CDR2 or CDR3. In: Journal of Molecular Biology. 2005 ; Vol. 346, No. 1. pp. 223-239.
@article{b0c03e3642284d1ea840bfe49683a27e,
title = "High-affinity, peptide-specific T cell receptors can be generated by mutations in CDR1, CDR2 or CDR3",
abstract = "The third complementarity-determining regions (CDR3s) of antibodies and T cell receptors (TCRs) have been shown to play a major role in antigen binding and specificity. Consistent with this notion, we demonstrated previously that high-affinity, peptide-specific TCRs could be generated in vitro by mutations in the CDR3α region of the 2C TCR. In contrast, it has been argued that CDR1 and CDR2 are involved to a greater extent than CDR3s in the process of MHC restriction, due to their engagement of MHC helices. Based on this premise, we initiated the present study to explore whether higher affinity TCRs generated through mutations in these CDRs or other regions would lead to significant reductions in peptide specificity (i.e. the result of greater binding energy gained through interactions with major histocompatibility complex (MHC) helices). Yeast-display technology and flow sorting were used to select high-affinity TCRs from libraries of CDR mutants or random mutants. High-affinity TCRs with mutations in the first residue of the Vα, CDR1, CDR2, or CDR3 were isolated. Unexpectedly, every TCR mutant, including those in CDR1 and CDR2, retained remarkable peptide specificity. Molecular modeling of various mutants suggested that such exquisite specificity may be due to: (1) enhanced electrostatic interactions with key peptide or MHC residues; or (2) stabilization of CDRs in specific conformations. The results indicate that the TCR is positioned so that virtually every CDR can contribute to the antigen-specificity of a T cell. The conserved diagonal docking of TCRs could thus orient each CDR loop to sense the peptide directly or indirectly through peptide-induced effects on the MHC.",
keywords = "T cell receptors, antigen recognition, protein engineering, yeast display",
author = "Chlewicki, {Lukasz K.} and Holler, {Phillip D.} and Monti, {Bridget C.} and Clutter, {Matthew R.} and Kranz, {David M.}",
year = "2005",
month = "2",
day = "11",
doi = "10.1016/j.jmb.2004.11.057",
language = "English (US)",
volume = "346",
pages = "223--239",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - High-affinity, peptide-specific T cell receptors can be generated by mutations in CDR1, CDR2 or CDR3

AU - Chlewicki, Lukasz K.

AU - Holler, Phillip D.

AU - Monti, Bridget C.

AU - Clutter, Matthew R.

AU - Kranz, David M.

PY - 2005/2/11

Y1 - 2005/2/11

N2 - The third complementarity-determining regions (CDR3s) of antibodies and T cell receptors (TCRs) have been shown to play a major role in antigen binding and specificity. Consistent with this notion, we demonstrated previously that high-affinity, peptide-specific TCRs could be generated in vitro by mutations in the CDR3α region of the 2C TCR. In contrast, it has been argued that CDR1 and CDR2 are involved to a greater extent than CDR3s in the process of MHC restriction, due to their engagement of MHC helices. Based on this premise, we initiated the present study to explore whether higher affinity TCRs generated through mutations in these CDRs or other regions would lead to significant reductions in peptide specificity (i.e. the result of greater binding energy gained through interactions with major histocompatibility complex (MHC) helices). Yeast-display technology and flow sorting were used to select high-affinity TCRs from libraries of CDR mutants or random mutants. High-affinity TCRs with mutations in the first residue of the Vα, CDR1, CDR2, or CDR3 were isolated. Unexpectedly, every TCR mutant, including those in CDR1 and CDR2, retained remarkable peptide specificity. Molecular modeling of various mutants suggested that such exquisite specificity may be due to: (1) enhanced electrostatic interactions with key peptide or MHC residues; or (2) stabilization of CDRs in specific conformations. The results indicate that the TCR is positioned so that virtually every CDR can contribute to the antigen-specificity of a T cell. The conserved diagonal docking of TCRs could thus orient each CDR loop to sense the peptide directly or indirectly through peptide-induced effects on the MHC.

AB - The third complementarity-determining regions (CDR3s) of antibodies and T cell receptors (TCRs) have been shown to play a major role in antigen binding and specificity. Consistent with this notion, we demonstrated previously that high-affinity, peptide-specific TCRs could be generated in vitro by mutations in the CDR3α region of the 2C TCR. In contrast, it has been argued that CDR1 and CDR2 are involved to a greater extent than CDR3s in the process of MHC restriction, due to their engagement of MHC helices. Based on this premise, we initiated the present study to explore whether higher affinity TCRs generated through mutations in these CDRs or other regions would lead to significant reductions in peptide specificity (i.e. the result of greater binding energy gained through interactions with major histocompatibility complex (MHC) helices). Yeast-display technology and flow sorting were used to select high-affinity TCRs from libraries of CDR mutants or random mutants. High-affinity TCRs with mutations in the first residue of the Vα, CDR1, CDR2, or CDR3 were isolated. Unexpectedly, every TCR mutant, including those in CDR1 and CDR2, retained remarkable peptide specificity. Molecular modeling of various mutants suggested that such exquisite specificity may be due to: (1) enhanced electrostatic interactions with key peptide or MHC residues; or (2) stabilization of CDRs in specific conformations. The results indicate that the TCR is positioned so that virtually every CDR can contribute to the antigen-specificity of a T cell. The conserved diagonal docking of TCRs could thus orient each CDR loop to sense the peptide directly or indirectly through peptide-induced effects on the MHC.

KW - T cell receptors

KW - antigen recognition

KW - protein engineering

KW - yeast display

UR - http://www.scopus.com/inward/record.url?scp=12344322695&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12344322695&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2004.11.057

DO - 10.1016/j.jmb.2004.11.057

M3 - Article

C2 - 15663940

AN - SCOPUS:12344322695

VL - 346

SP - 223

EP - 239

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 1

ER -