Crystallization and preliminary X-ray structural studies of a high-affinity CD8alphaalpha co-receptor to pMHC

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Mar 1;61(Pt 3):285-7. doi: 10.1107/S1744309105002988. Epub 2005 Feb 12.

Abstract

The class I CD8 positive T-cell response is involved in a number of conditions in which artificial down-regulation and control would be therapeutically beneficial. Such conditions include a number of autoimmune diseases and graft rejection in transplant patients. Although the CD8 T-cell response is dominated by the TCR-pMHC interaction, activation of T cells is in most cases also dependent on a number of associated signalling molecules. Previous work has demonstrated the ability of one such molecule (CD8) to act as an antagonist to T-cell activation if added in soluble form. Therefore, a high-affinity mutant CD8 (haCD8) has been developed with the aim of developing a therapeutic immunosuppressor. In order to fully understand the nature of the haCD8 interaction, this protein was crystallized using the sitting-drop vapour-diffusion method. Single haCD8 crystals were cryocooled and used for data collection. These crystals belonged to space group P6(4)22 (assumed by similarity to the wild type), with unit-cell parameters a = 101.08, c = 56.54 A. VM calculations indicated one molecule per asymmetric unit. A 2 A data set was collected and the structure is currently being determined using molecular replacement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD8-Positive T-Lymphocytes / immunology
  • Cloning, Molecular
  • Crystallization
  • Humans
  • Major Histocompatibility Complex*
  • Protein Conformation
  • Protein Denaturation
  • Protein Folding
  • Receptors, Antigen, T-Cell / chemistry*
  • Receptors, Antigen, T-Cell / isolation & purification
  • Receptors, Antigen, T-Cell / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • CD8 receptor
  • Receptors, Antigen, T-Cell
  • Recombinant Proteins