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Recombinant Amyloid Beta-peptide Production by Coexpression with an Affibody Ligand by Bertil Macao; Wolfgang Hoyer; Anders Sandberg; Ann-Christin Brorsson; Christopher M Dobson; Torleif Härd is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Background Oligomeric and fibrillar aggregates of the amyloid β-peptide (Aβ) have been implicated in the pathogenesis of Alzheimer's disease (AD). The characterization of Aβ assemblies is essential for the elucidation of the mechanisms of Aβ neurotoxicity, but requires large quantities of pure peptide. Here we describe a novel approach to the recombinant production of Aβ. The method is based on the coexpression of the affibody protein Z~Aβ3~, a selected affinity ligand derived from the Z domain three-helix bundle scaffold. Z~Aβ3~ binds to the amyloidogenic central and C-terminal part of Aβ with nanomolar affinity and consequently inhibits aggregation. ## Results Coexpression of Z~Aβ3~ affords the overexpression of both major Aβ isoforms, Aβ(1–40) and Aβ(1–42), yielding 4 or 3 mg, respectively, of pure ^15^N-labeled peptide per liter of culture. The method does not rely on a protein-fusion or -tag and thus does not require a cleavage reaction. The purified peptides were characterized by NMR, circular dichroism, SDS-PAGE and size exclusion chromatography, and their aggregation propensities were assessed by thioflavin T fluorescence and electron microscopy. The data coincide with t

Who reads Recombinant Amyloid Beta-peptide Production by Coexpression with an Affibody Ligand?

Author

Bertil Macao; Wolfgang Hoyer; Anders Sandberg; Ann-Christin Brorsson; Christopher M Dobson; Torleif Härd

Publisher

BioMed Central; Springer (Biomed Central Ltd.); [London]: BioMed Central, 2001-; Springer Science and Business Media LLC (ISSN 1472-6750)

Published

2008

Language

EN

ISBN

9781472675088

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)