Design, engineering and production of functional single-chain T cell receptor ligands

G. G. Burrows, J. W. Chang, H. P. Bächinger, D. N. Bourdette, H. Offner, A. A. Vandenbark

Research output: Contribution to journalArticlepeer-review

56 Scopus citations


Major histocompatibility complex (MHC) class II molecules are membrane-anchored heterodimers on the surface of antigen presenting cells (APCs) that bind the T cell receptor, initiating a cascade of interactions that results in antigen-specific activation of clonal populations of T cells. The peptide binding/T cell recognition domains of rat MHC class II (alpha-1 and beta-1 domains) were expressed as a single exon for structural and functional characterization. These recombinant single-chain T cell receptor ligands (termed 'β1α1' molecules) of approximately 200 amino acid residues were designed using the structural backbone of MHC class II molecules as template, and have been produced in Escherichia coli with and without N-terminal extensions containing antigenic peptides. Structural characterization using circular dichroism predicted that these molecules retained the antiparallel β-sheet platform and antiparallel α-helices observed in the native MHC class II heterodimer. The proteins exhibited a cooperative two-state thermal folding-unfolding transition. β1α1 molecules with a covalently linked MBP-72-89 peptide showed increased stability to thermal unfolding relative to the empty β1α1 molecules. This new class of small soluble polypeptide provides a template for designing and refining human homologues useful in detecting and regulating pathogenic T cells.

Original languageEnglish (US)
Pages (from-to)771-778
Number of pages8
JournalProtein Engineering
Issue number9
StatePublished - 1999


  • Drug design
  • Immunotherapy
  • MHC class II
  • TCR ligand
  • β1α1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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