Backbone resonance assignment of N15, N30 and D10 T cell receptor β subunits.
The αβT Cell receptor (TCR) governs T cell immunity through its interaction with peptide bound to major histocompatibility complex molecules (pMHC). Previously, soluble ectodomain constructs have been used to elucidate the binding mode of the TCR for the MHC. However, the full heterodimeric αβTCR has proven difficult to produce reproducibly in recombinant systems to the extent seen in the routine production of novel antibodies. Particularly, the route of production in E. coli, which is most convenient for isotopic labeling of proteins, is challenging for a wide range of αβTCR, including N15αβ, N30αβ, but not D10αβ. With the aim of understanding the TCR-pMHC interaction through the use of dynamic binding measurements, we set out to produce TCRβ subunits with which we could investigate binding with pMHC. The TCRβ constructs are more readily produced and refolded than their αβ counterparts and have proven to be an effective model of preTCR in pMHC binding studies. As a first step towards characterizing potential interactions with protein ligands, we have assigned the backbone resonances of three TCRβ subunits, N15β, N30β and D10β.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA.
Robert J
RJ
Mallis
Laboratory of Immunobiology, Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02115, USA.|Department of Medicine, Harvard Medical School, Boston, MA, 02115, USA.
Ellis L
EL
Reinherz
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA.
Gerhard
G
Wagner
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA. hari_arthanari@hms.harvard.edu.
Haribabu
H
Arthanari
D000595:Amino Acid Sequence; D019906:Nuclear Magnetic Resonance, Biomolecular; D000072417:Protein Domains; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry*; D016415:Sequence Alignment
D021122:Protein Subunits; D016693:Receptors, Antigen, T-Cell, alpha-beta
Immunology; MHC; Protein refolding; T-cell receptor; TCR