High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain

Sci Rep. 2020 Jan 14;10(1):235. doi: 10.1038/s41598-019-57143-x.

Abstract

During storage in the silk gland, the N-terminal domain (NT) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that NTs from different spidroins have co-evolved with their respective repeat region, and now use an NT that is distantly related to previously used NTs, for efficient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer's disease. A designed variant of NT from Nephila clavipes flagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of different human Aβ variants as a solubility tag. This tool enables efficient production of target peptides also in minimal medium and gives up to 10 times more isotope-labeled monomeric Aβ peptides per liter bacterial culture than previously reported.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amyloid beta-Peptides / metabolism*
  • Animals
  • Fibroins / chemistry*
  • Fibroins / metabolism*
  • Models, Molecular
  • Protein Domains

Substances

  • Amyloid beta-Peptides
  • Fibroins