Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions
File(s)CD4 pnas ms and si rev.pdf (1.56 MB)
Accepted version
Author(s)
Type
Journal Article
Abstract
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/μm2. 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.
Date Issued
2016-05-17
Date Acceptance
2016-03-23
Citation
Proceedings of the National Academy of Sciences of the United States of America, 2016, 113 (20), pp.5682-5687
ISSN
1091-6490
Publisher
National Academy of Sciences
Start Page
5682
End Page
5687
Journal / Book Title
Proceedings of the National Academy of Sciences of the United States of America
Volume
113
Issue
20
Copyright Statement
© 2016 National Academy of Sciences.
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000375977600057&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
Protein interactions
TCR phosphorylation
Adhesion
T-cell activation
Binding equilibrium
Kinetics
T-cell-receptor
Conctact-area FRAP
Crystal-structure
Antigen discrimination
Immunological synapse
Fluorescence recovery
Peptide-MHC
Molecules
Binding equilibrium and kinetics
MD Multidisciplinary
Publication Status
Published
Date Publish Online
2016-04-25