Regulation of T Cell Receptor Activation by Dynamic Membrane Binding of the CD3ɛ Cytoplasmic Tyrosine-Based Motif

Many immune system receptors signal through cytoplasmic tyrosine-based motifs (ITAMs), but how receptor ligation results in ITAM phosphorylation remains unknown. Live-cell imaging studies showed a close interaction of the CD3epsilon cytoplasmic domain of the T cell receptor (TCR) with the plasma membrane through fluorescence resonance energy transfer between a C-terminal fluorescent protein and a membrane fluorophore. Electrostatic interactions between basic CD3epsilon residues and acidic phospholipids enriched in the inner leaflet of the plasma membrane were required for binding. The nuclear magnetic resonance structure of the lipid-bound state of this cytoplasmic domain revealed deep insertion of the two key tyrosines into the hydrophobic core of the lipid bilayer. Receptor ligation thus needs to result in unbinding of the CD3epsilon ITAM from the membrane to render these tyrosines accessible to Src kinases. Sequestration of key tyrosines into the lipid bilayer represents a previously unrecognized mechanism for control of receptor activation.

[1]  Martin Wiedmann,et al.  The Organizing Principle in the Formation of the T Cell Receptor-CD3 Complex , 2002, Cell.

[2]  Zhengyu Ma,et al.  Surface-Anchored Monomeric Agonist pMHCs Alone Trigger TCR with High Sensitivity , 2008, PLoS biology.

[3]  Balbino Alarcón,et al.  Recruitment of Nck by CD3ϵ Reveals a Ligand-Induced Conformational Change Essential for T Cell Receptor Signaling and Synapse Formation , 2002, Cell.

[4]  Mark M. Davis,et al.  Deconstructing the form and function of the TCR/CD3 complex. , 2006, Immunity.

[5]  Robert E Campbell,et al.  Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging. , 2006, The Biochemical journal.

[6]  Mark Philips,et al.  Receptor Activation Alters Inner Surface Potential During Phagocytosis , 2006, Science.

[7]  S. McLaughlin,et al.  Binding of Peptides with Basic and Aromatic Residues to Bilayer Membranes , 2003, Journal of Biological Chemistry.

[8]  M. Reth.  Antigen receptor tail clue , 1989, Nature.

[9]  E. Reinherz,et al.  Mechanisms Contributing to T Cell Receptor Signaling and Assembly Revealed by the Solution Structure of an Ectodomain Fragment of the CD3ϵγ Heterodimer , 2001, Cell.

[10]  V. Uversky,et al.  Lipid-binding activity of intrinsically unstructured cytoplasmic domains of multichain immune recognition receptor signaling subunits. , 2006, Biochemistry.

[11]  Charles D Schwieters,et al.  The Xplor-NIH NMR molecular structure determination package. , 2003, Journal of magnetic resonance.

[12]  A. Bax,et al.  Micelle-induced curvature in a water-insoluble HIV-1 Env peptide revealed by NMR dipolar coupling measurement in stretched polyacrylamide gel. , 2002, Journal of the American Chemical Society.

[13]  Ellen R. Laird,et al.  Molecular basis for interaction of the protein tyrosine kinase ZAP-70 with the T-cell receptor , 2007, Nature.

[14]  P. Devaux,et al.  Static and dynamic lipid asymmetry in cell membranes. , 1991, Biochemistry.

[15]  Dan R. Littman,et al.  Signal transduction by lymphocyte antigen receptors , 1994, Cell.

[16]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[17]  A. Kenworthy,et al.  Imaging protein-protein interactions using fluorescence resonance energy transfer microscopy. , 2001, Methods.

[18]  S. McLaughlin,et al.  The Effector Domain of Myristoylated Alanine-rich C Kinase Substrate Binds Strongly to Phosphatidylinositol 4,5-Bisphosphate* , 2001, The Journal of Biological Chemistry.

[19]  R. Prosser,et al.  Current applications of bicelles in NMR studies of membrane-associated amphiphiles and proteins. , 2006, Biochemistry.

[20]  P. Gimotty,et al.  An Immunoreceptor Tyrosine Activation Motif in the Mouse Mammary Tumor Virus Envelope Protein Plays a Role in Virus-Induced Mammary Tumors , 2006, Journal of Virology.

[21]  D. Aivazian,et al.  Phosphorylation of T cell receptor ζ is regulated by a lipid dependent folding transition , 2000, Nature Structural Biology.

[22]  M. Freeman,et al.  Draper-dependent glial phagocytic activity is mediated by Src and Syk family kinase signalling , 2008, Nature.

[23]  E. Komives,et al.  Structural evaluation of phospholipid bicelles for solution-state studies of membrane-associated biomolecules. , 2001, Biophysical journal.

[24]  S. Caughman,et al.  T-cell antigen receptors. , 1990, Dermatologic clinics.

[25]  S H White,et al.  Structure of a fluid dioleoylphosphatidylcholine bilayer determined by joint refinement of x-ray and neutron diffraction data. III. Complete structure. , 1992, Biophysical journal.

[26]  John Kuriyan,et al.  Structural Basis for the Inhibition of Tyrosine Kinase Activity of ZAP-70 , 2007, Cell.

[27]  E. Reinherz,et al.  Mechanisms contributing to T cell receptor signaling and assembly revealed by the solution structure of an ectodomain fragment of the CD3 epsilon gamma heterodimer. , 2001, Cell.

[28]  Eric R. Prossnitz,et al.  FRET Detection of Cellular α4-Integrin Conformational Activation , 2003 .

[29]  R. Longnecker,et al.  Syk Tyrosine Kinase Mediates Epstein-Barr Virus Latent Membrane Protein 2A-induced Cell Migration in Epithelial Cells* , 2006, Journal of Biological Chemistry.

[30]  P. Allen,et al.  Fidelity of T cell activation through multistep T cell receptor zeta phosphorylation. , 1998, Science.

[31]  T. Meyer,et al.  PI(3,4,5)P3 and PI(4,5)P2 Lipids Target Proteins with Polybasic Clusters to the Plasma Membrane , 2006, Science.

[32]  T. Jin,et al.  Ligand-Induced Partitioning of Human CXCR1 Chemokine Receptors with Lipid Raft Microenvironments Facilitates G-Protein-Dependent Signaling , 2005, Molecular and Cellular Biology.

[33]  K. Wucherpfennig,et al.  Common themes in the assembly and architecture of activating immune receptors , 2007, Nature Reviews Immunology.

[34]  A. Chigaev,et al.  FRET detection of cellular alpha4-integrin conformational activation. , 2003, Biophysical journal.

[35]  Marie Malissen,et al.  The proline-rich sequence of CD3ε controls T cell antigen receptor expression on and signaling potency in preselection CD4+CD8+ thymocytes , 2008, Nature Immunology.

[36]  Mark M. Davis,et al.  T cell antigen receptor , 1986 .

[37]  F W McLafferty,et al.  Quantitative analysis of phospholipids in functionally important membrane domains from RBL-2H3 mast cells using tandem high-resolution mass spectrometry. , 1999, Biochemistry.