Computational protocol: Combining intracellular selection with protein-fragment complementation to derive Aβ interacting peptides

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Protocol publication

[…] Peptides KAT, L2P1a, L2P1b, L2P2a, L2P2b and a positive control from the literature, iAβ5 (), were obtained by Peptide Protein Research (Fareham, UK) as pure lyophilised peptides and weighed using an analytical balance. Stock solutions of 1 mM concentration were subsequently dissolved in ultrapure water. At this concentration of a 2–200-fold excess of that used in inhibition and reversal experiments, no aggregation or precipitate was observed. In addition bioinformatics tools (e.g. Waltz (), Amylpred (), Pasta (), Zyggregator () and Tango ()) did not predict any of the peptides to contain amyloidogenic sequences or aggregate in isolation. Finally, dye-binding experiments demonstrate that these sequences do not bind thioflavin-T (ThT) and form random-coil like species in isolation using circular dichroism (CD) (see Supplementary information).See Supplementary information for additional methods. […]

Pipeline specifications

Software tools AMYLPRED, PASTA, Zyggregator, TANGO
Application Protein physicochemical analysis
Organisms Escherichia coli
Diseases Alzheimer Disease, Amyloidosis