3s4s Citations

Affinity maturation of human CD4 by yeast surface display and crystal structure of a CD4-HLA-DR1 complex.

Proc. Natl. Acad. Sci. U.S.A. 108 15960-5 (2011)
Cited: 14 times
EuropePMC logo PMID: 21900604


Helper T-cell activation generally requires the coreceptor CD4, which binds MHC class II molecules. A remarkable feature of the CD4-MHC class II interaction is its exceptionally low affinity, which ranges from K(D) = ∼200 μM to >2 mM. Investigating the biological role of the much lower affinity of this interaction than those of other cell-cell recognition molecules will require CD4 mutants with enhanced binding to MHC class II for testing in models of T-cell development. To this end, we used in vitro-directed evolution to increase the affinity of human CD4 for HLA-DR1. A mutant CD4 library was displayed on the surface of yeast and selected using HLA-DR1 tetramers or monomers, resulting in isolation of a CD4 clone containing 11 mutations. Reversion mutagenesis showed that most of the affinity increase derived from just two substitutions, Gln40Tyr and Thr45Trp. A CD4 variant bearing these mutations bound HLA-DR1 with K(D) = 8.8 μM, compared with >400 μM for wild-type CD4. To understand the basis for improved affinity, we determined the structure of this CD4 variant in complex with HLA-DR1 to 2.4 Å resolution. The structure provides an atomic-level description of the CD4-binding site on MHC class II and reveals how CD4 recognizes highly polymorphic HLA-DR, -DP, and -DQ molecules by targeting invariant residues in their α2 and β2 domains. In addition, the CD4 mutants reported here constitute unique tools for probing the influence of CD4 affinity on T-cell activation and development.

Articles - 3s4s mentioned but not cited (1)

Reviews citing this publication (7)

  1. The versatility of the αβ T-cell antigen receptor. Bhati M, Cole DK, McCluskey J, Sewell AK, Rossjohn J. Protein Sci. 23 260-272 (2014)
  2. Strict Major Histocompatibility Complex Molecule Class-Specific Binding by Co-Receptors Enforces MHC-Restricted αβ TCR Recognition during T Lineage Subset Commitment. Li XL, Teng MK, Reinherz EL, Wang JH. Front Immunol 4 383 (2013)
  3. Selection of antibodies from synthetic antibody libraries. Harel Inbar N, Benhar I. Arch. Biochem. Biophys. 526 87-98 (2012)
  4. Engineering antibodies by yeast display. Boder ET, Raeeszadeh-Sarmazdeh M, Price JV. Arch. Biochem. Biophys. 526 99-106 (2012)
  5. Structural basis for self-recognition by autoimmune T-cell receptors. Yin Y, Li Y, Mariuzza RA. Immunol. Rev. 250 32-48 (2012)
  6. Diversity-oriented approaches for interrogating T-cell receptor repertoire, ligand recognition, and function. Birnbaum ME, Dong S, Garcia KC. Immunol. Rev. 250 82-101 (2012)
  7. The structural basis of αβ T-lineage immune recognition: TCR docking topologies, mechanotransduction, and co-receptor function. Wang JH, Reinherz EL. Immunol. Rev. 250 102-119 (2012)

Articles citing this publication (6)

  1. Crystal structure of a complete ternary complex of T-cell receptor, peptide-MHC, and CD4. Yin Y, Wang XX, Mariuzza RA. Proc. Natl. Acad. Sci. U.S.A. 109 5405-5410 (2012)
  2. Comprehensive analysis of MHC class II genes in teleost fish genomes reveals dispensability of the peptide-loading DM system in a large part of vertebrates. Dijkstra JM, Grimholt U, Leong J, Koop BF, Hashimoto K. BMC Evol. Biol. 13 260 (2013)
  3. Conformation-dependent epitopes recognized by prion protein antibodies probed using mutational scanning and deep sequencing. Doolan KM, Colby DW. J. Mol. Biol. 427 328-340 (2015)
  4. Structural and biophysical insights into the role of CD4 and CD8 in T cell activation. Li Y, Yin Y, Mariuzza RA. Front Immunol 4 206 (2013)
  5. Islet-Derived CD4 T Cells Targeting Proinsulin in Human Autoimmune Diabetes. Michels AW, Landry LG, McDaniel KA, Yu L, Campbell-Thompson M, Kwok WW, Jones KL, Gottlieb PA, Kappler JW, Tang Q, Roep BO, Atkinson MA, Mathews CE, Nakayama M. Diabetes 66 722-734 (2017)
  6. Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions. Jönsson P, Southcombe JH, Santos AM, Huo J, Fernandes RA, McColl J, Lever M, Evans EJ, Hudson A, Chang VT, Hanke T, Godkin A, Dunne PD, Horrocks MH, Palayret M, Screaton GR, Petersen J, Rossjohn J, Fugger L, Dushek O, Xu XN, Davis SJ, Klenerman D. Proc. Natl. Acad. Sci. U.S.A. 113 5682-5687 (2016)