The role of interchain disulfide bond in a recombinant human interleukin-17A variant

Cytokine. 2014 Feb;65(2):167-74. doi: 10.1016/j.cyto.2013.11.007. Epub 2013 Dec 15.

Abstract

Interleukin-17A (IL-17A) is the prototype of IL-17 family and has been implicated in the pathogenesis of a variety of autoimmune diseases. Therefore its structural and functional properties are of great medical interest. During our research on a recombinant human IL-17A (rhIL-17A) variant, four isoforms were obtained when it was refolded. While isoforms 1 and 2 represented non-covalent dimers, isoforms 3 and 4 were determined to be covalent dimers. All four isoforms were structurally similar by Circular Dichroism and fluorescence spectroscopy studies, but differential scanning calorimetry demonstrated thermal stability in the order of isoform 1=isoform 2<isoform 4<isoform 3. In addition, compared to covalent dimers (isoform 3 and 4), the non-covalent dimers (isoforms 1 and 2) are slightly less active in a receptor-binding assay but at least 5-fold less active in a cell-based assay.

Keywords: Activity; CD; CHCA; Circular Dichroism; DSC; Disulfide bond; HPBA; IL-17A; MMmeas; MMtheo; PrOH; Refolding; alpha-cyano-4-hydroxycinnamic acid; differential scanning calorimetry; heptafluoro-butyric anhydride; measured molar mass; propanol; theoretical molar mass.

MeSH terms

  • Amino Acid Sequence
  • Calorimetry, Differential Scanning
  • Circular Dichroism
  • Disulfides / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Interleukin-17 / chemistry
  • Interleukin-17 / metabolism*
  • Mass Spectrometry
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / metabolism
  • Protein Isoforms / metabolism
  • Protein Multimerization
  • Protein Refolding
  • Recombinant Proteins / metabolism*
  • Solutions
  • Spectrometry, Fluorescence

Substances

  • Disulfides
  • IL17A protein, human
  • Interleukin-17
  • Peptides
  • Protein Isoforms
  • Recombinant Proteins
  • Solutions