Characterization of a Putative Receptor Binding Surface on Skint-1, a Critical Determinant of Dendritic Epidermal T Cell Selection

J Biol Chem. 2016 Apr 22;291(17):9310-21. doi: 10.1074/jbc.M116.722066. Epub 2016 Feb 25.

Abstract

Dendritic epidermal T cells (DETC) form a skin-resident γδ T cell population that makes key contributions to cutaneous immune stress surveillance, including non-redundant contributions to protection from cutaneous carcinogens. How DETC become uniquely associated with the epidermis was in large part solved by the identification of Skint-1, the prototypic member of a novel B7-related multigene family. Expressed only by thymic epithelial cells and epidermal keratinocytes, Skint-1 drives specifically the development of DETC progenitors, making it the first clear candidate for a selecting ligand for non-MHC/CD1-restricted T cells. However, the molecular mechanisms underpinning Skint-1 activity are unresolved. Here, we provide evidence that DETC selection requires Skint-1 expression on the surface of thymic epithelial cells, and depends upon specific residues on the CDR3-like loop within the membrane-distal variable domain of Skint-1 (Skint-1 DV). Nuclear magnetic resonance of Skint-1 DV revealed a core tertiary structure conserved across the Skint family, but a highly distinct surface charge distribution, possibly explaining its unique function. Crucially, the CDR3-like loop formed an electrostatically distinct surface, featuring key charged and hydrophobic solvent-exposed residues, at the membrane-distal tip of DV. These results provide the first structural insights into the Skint family, identifying a putative receptor binding surface that directly implicates Skint-1 in receptor-ligand interactions crucial for DETC selection.

Keywords: T-cell receptor (TCR); immunology; lymphocyte; nuclear magnetic resonance (NMR); stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Epidermis / immunology*
  • Epithelial Cells / immunology*
  • Humans
  • Immunoglobulins / chemistry*
  • Immunoglobulins / immunology*
  • Mice
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Receptors, Antigen, T-Cell, gamma-delta / immunology*
  • T-Lymphocytes / immunology*

Substances

  • Immunoglobulins
  • Receptors, Antigen, T-Cell, gamma-delta
  • Skint1 protein, mouse

Associated data

  • PDB/1PY9
  • PDB/2N4I
  • PDB/3FN3
  • PDB/4F8T
  • PDB/4F9P
  • PDB/4HH8
  • PDB/4K55