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

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

Author information

1
Department of Neurobiology, Care Sciences and Society, Center for Alzheimer Research, Division of Neurogeriatrics, Karolinska Institutet, 141 52, Huddinge, Sweden.
2
Department of Physical Organic Chemistry, Latvian Institute of Organic Synthesis, Riga, LV-1006, Latvia.
3
Department of Molecular Tumor and Cell Biology, Karolinska Institutet, 171 65, Solna, Sweden.
4
Department of Pathology, University of Oslo/Oslo University Hospital, N-0424, Oslo, Norway.
5
LIED, University of Lübeck, D-23538, Lübeck, Germany.
6
Department of Pharmacology, Medical Faculty, University of Latvia, Riga, LV-1004, Latvia.
7
Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, 750 07, Uppsala, Sweden.
8
Department of Neurobiology, Care Sciences and Society, Center for Alzheimer Research, Division of Neurogeriatrics, Karolinska Institutet, 141 52, Huddinge, Sweden. henrik.biverstal@ki.se.
9
Department of Physical Organic Chemistry, Latvian Institute of Organic Synthesis, Riga, LV-1006, Latvia. henrik.biverstal@ki.se.

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.

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