Abstract
T cell receptor (TCR) recognition of peptide-MHC (pMHC) is central to the cellular immune response. A large database of TCR-pMHC structures is needed to reveal general structural principles, such as whether the repertoire of TCR/MHC docking modes is dictated by a "recognition code" between conserved elements of the TCR and MHC genes. Although ≈17 cocrystal structures of unique TCR-pMHC complexes have been determined, cocrystallization of soluble TCR and pMHC remains a major technical obstacle in the field. Here we demonstrate a strategy, based on NMR chemical shift mapping, that permits rapid and reliable analysis of the solution footprint made by a TCR when binding onto the pMHC surface. We mapped the 2C TCR binding interaction with its allogeneic ligand H-2Ld-QL9 and identified a group of NMR-shifted residues that delineated a clear surface of the MHC that we defined as the TCR footprint. We subsequently found that the docking footprint described by NMR shifts was highly accurate compared with a recently determined high-resolution crystal structure of the same complex. The same NMR footprint analysis was done on a high-affinity mutant of the TCR. The current work serves as a foundation to explore the molecular dynamics of pMHC complexes and to rapidly determine the footprints of many Ld-specific TCRs.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 13080-13085 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 104 |
| Issue number | 32 |
| DOIs | |
| State | Published - Aug 7 2007 |
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Keywords
- Cellular immunity
- Chemical shift mapping
- Dynamics
- NMR
- Protein-protein interaction
ASJC Scopus subject areas
- General
Cite this
Solution mapping of T cell receptor docking footprints on peptide-MHC. / Varani, Luca; Bankovich, Alexander J.; Liu, Corey W.; Colf, Leremy A.; Jones, Lindsay L.; Kranz, David M.; Puglisi, Joseph D.; Garcia, K. Christopher.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 32, 07.08.2007, p. 13080-13085.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Solution mapping of T cell receptor docking footprints on peptide-MHC
AU - Varani, Luca
AU - Bankovich, Alexander J.
AU - Liu, Corey W.
AU - Colf, Leremy A.
AU - Jones, Lindsay L.
AU - Kranz, David M.
AU - Puglisi, Joseph D.
AU - Garcia, K. Christopher
PY - 2007/8/7
Y1 - 2007/8/7
N2 - T cell receptor (TCR) recognition of peptide-MHC (pMHC) is central to the cellular immune response. A large database of TCR-pMHC structures is needed to reveal general structural principles, such as whether the repertoire of TCR/MHC docking modes is dictated by a "recognition code" between conserved elements of the TCR and MHC genes. Although ≈17 cocrystal structures of unique TCR-pMHC complexes have been determined, cocrystallization of soluble TCR and pMHC remains a major technical obstacle in the field. Here we demonstrate a strategy, based on NMR chemical shift mapping, that permits rapid and reliable analysis of the solution footprint made by a TCR when binding onto the pMHC surface. We mapped the 2C TCR binding interaction with its allogeneic ligand H-2Ld-QL9 and identified a group of NMR-shifted residues that delineated a clear surface of the MHC that we defined as the TCR footprint. We subsequently found that the docking footprint described by NMR shifts was highly accurate compared with a recently determined high-resolution crystal structure of the same complex. The same NMR footprint analysis was done on a high-affinity mutant of the TCR. The current work serves as a foundation to explore the molecular dynamics of pMHC complexes and to rapidly determine the footprints of many Ld-specific TCRs.
AB - T cell receptor (TCR) recognition of peptide-MHC (pMHC) is central to the cellular immune response. A large database of TCR-pMHC structures is needed to reveal general structural principles, such as whether the repertoire of TCR/MHC docking modes is dictated by a "recognition code" between conserved elements of the TCR and MHC genes. Although ≈17 cocrystal structures of unique TCR-pMHC complexes have been determined, cocrystallization of soluble TCR and pMHC remains a major technical obstacle in the field. Here we demonstrate a strategy, based on NMR chemical shift mapping, that permits rapid and reliable analysis of the solution footprint made by a TCR when binding onto the pMHC surface. We mapped the 2C TCR binding interaction with its allogeneic ligand H-2Ld-QL9 and identified a group of NMR-shifted residues that delineated a clear surface of the MHC that we defined as the TCR footprint. We subsequently found that the docking footprint described by NMR shifts was highly accurate compared with a recently determined high-resolution crystal structure of the same complex. The same NMR footprint analysis was done on a high-affinity mutant of the TCR. The current work serves as a foundation to explore the molecular dynamics of pMHC complexes and to rapidly determine the footprints of many Ld-specific TCRs.
KW - Cellular immunity
KW - Chemical shift mapping
KW - Dynamics
KW - NMR
KW - Protein-protein interaction
UR - http://www.scopus.com/inward/record.url?scp=34548736431&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548736431&partnerID=8YFLogxK
U2 - 10.1073/pnas.0703702104
DO - 10.1073/pnas.0703702104
M3 - Article
C2 - 17670943
AN - SCOPUS:34548736431
VL - 104
SP - 13080
EP - 13085
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 32
ER -