| Autor: |
Jönsson, Peter, Southcombe, Jennifer, Santos, Ana Mafalda, Huo, Jiandong, Fernandes, Ricardo A, McColl, James, Lever, Melissa, Evans, Edward J, Hudson, Alexander, Chang, Veronica T, Hanke, Tomáš, Godkin, Andrew, Dunne, Paul D, Horrocks, Mathew H, Palayret, Matthieu, Screaton, Gavin R, Petersen, Jan, Rossjohn, Jamie, Fugger, Lars, Dushek, Omer, Xu, Xiaoning, Davis, Simon J, Klenerman, David |
| Přispěvatelé: |
Klenerman, David [0000-0001-7116-6954], Apollo - University of Cambridge Repository |
| Jazyk: |
angličtina |
| Rok vydání: |
2016 |
| Předmět: |
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| Popis: |
The αβ T-cell co-receptor CD4 enhances immune responses more than one 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 co-receptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in two dimensions (2D) using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D dissociation constant, Kd, of ~5000 molecules/μm2. This value is 2-3 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 three-fold 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 appears to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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