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.
| Original language | English |
|---|---|
| Article number | 235 |
| Journal | Scientific Reports |
| Volume | 10 |
| Issue number | 1 |
| ISSN | 2045-2322 |
| DOIs | |
| Publication status | Published - 01.12.2020 |
Funding
This study was supported by the Magnus Bergvall foundation (A.A., H.B.), FLPP/Latvia lzp-2018/1-0275 project support (H.B., J.P.), Swedish Research Council (A.R., J.J.), Center for Innovative Medicine (CIMED) (A.R. J.J.), FORMAS (A.R.), Vinnova (J.J.), Olle Engkvists Foundation (G.C.), Swedish Alzheimer foundation (G.C.), Åhlen Foundation (A.A., G.C., H.B.), Stiftelsen för Gamla Tjänarinnor (A.A., H.B., G.C.), P. & A. Hedlunds Stiftelse (G.C.), Instruct R&D pilot project grant APPID 272 (A.A.), Loo and Hans Osterman Foundation (A.A., G.C.), Geriatric Diseases Foundation at Karolinska Institutet (A.A., G.C.), Norsk forskningsrådet 251290, 260786 (J.P.), JPco-fuND/EU PROP-AD 643417/Horizon 2020/EC 643417 (H.B., J.P.).
Research Areas and Centers
- Academic Focus: Center for Infection and Inflammation Research (ZIEL)
