Abstract

Spiders have evolved proteins that can be kept in a highly concentrated soluble form in the silk gland yet rapidly assemble into stable silk fibers under certain environmental conditions. The transition between soluble and fibrillar states is partly regulated by the pH-sensitive N-terminal (NT) domain which has emerged as nature’s own solubility-enhancing domain. NT has an inherent capacity to keep the silk proteins’ partly hydrophobic and very aggregation-prone regions from premature fibrillation in spite of storage at enormous concentrations. The genetically engineered double-mutant NT* shows increased solubility and stability and has arisen as a powerful tool for the production of aggregation-prone as well as other recombinant proteins. Here we describe a robust and highly efficient protocol for improved soluble expression of peptides and proteins by fusion to the NT* tag.

Keywords

Aggregation-prone; Amyloid; NT domain; Recombinant production; Solubility tag; Spider silk protein; Surfactant protein; Transmembrane peptide

Published in

Methods in Molecular Biology
2022, Volume: 2406, number: 2406, pages: 113-130 Title: Insoluble Proteins : Methods and Protocols
ISBN: 978-1-0716-1858-5, eISBN: 978-1-0716-1859-2
Publisher: Springer

SLU Authors

• Rising, Anna

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    
  • Karolinska Institute

UKÄ Subject classification

Biochemistry and Molecular Biology


Publication identifier

DOI: https://doi.org/10.1007/978-1-0716-1859-2_6

Permanent link to this page (URI)

https://res.slu.se/id/publ/129699