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The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.

Original publication

DOI

10.1073/pnas.1513918113

Type

Journal article

Journal

Proc Natl Acad Sci U S A

Publication Date

17/05/2016

Volume

113

Pages

5682 - 5687

Keywords

T-cell activation, TCR phosphorylation, adhesion, binding equilibrium and kinetics, protein interactions, Binding Sites, CD4 Antigens, HEK293 Cells, HLA-A24 Antigen, HLA-DRB1 Chains, Humans, Maltose-Binding Proteins, Models, Molecular, Phosphorylation, Protein Binding, Protein Interaction Domains and Motifs, Protein Interaction Mapping, Protein Processing, Post-Translational, Protein Stability, Surface Plasmon Resonance