High yield production of amyloid-ß peptide enabled by a customized spider silk domain
2020
Abelein, Axel | Chen, Gefei | Kitoka, Kristīne | Aleksis, Rihards | Oleskovs, Filips | Sarr, Médoune | Landreh, Michael | Pahnke, Jens | Nordling, Kerstin | Kronqvist, Nina | Jaudzems, Kristaps | Rising, Anna | Johansson, Jan | Biverstal, Henrik
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 efcient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer’s disease. A designed variant of NT from Nephila clavipes fagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of diferent 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.
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