title: Tag-Free Semi-Synthesis of the Tau Protein.
abstract: Expressed protein ligation (EPL) is a valuable tool to study site-specific functionalities on proteins such as posttranslational modifications. The purification of such ligation products from EPL mixtures can be cumbersome due to a small size difference between the expressed protein portion and the desired ligated protein. Therefore, affinity tags are often required, which remain on the protein after purification. Herein, we present an efficient protocol to install a photocleavable biotin building block on synthetic C-terminal tau[390-441] and describe its use for purification of full-length semi-synthetic tau[1-441].
authors.affiliation: Leibniz-Institute for Molecular Pharmacology (FMP), Robert-Roessle-Strasse 10, 13125, Berlin, Germany. reimann@fmp-berlin.de.|Freie Universitaet Berlin, Institute for Chemistry und Biochemistry, Takustrasse 3, 14195, Berlin, Germany. reimann@fmp-berlin.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Reimann
authors.affiliation: Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France.|Research Federation FraBio 3688, Villeneuve d'Ascq, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Smet-Nocca
authors.affiliation: Leibniz-Institute for Molecular Pharmacology (FMP), Robert-Roessle-Strasse 10, 13125, Berlin, Germany.|Department of Chemistry, Humboldt Universitaet zu Berlin, Brook-Taylor-Strasse 2, 12489, Berlin, Germany.
authors.forename: Christian P R
authors.initials: CP
authors.lastname: Hackenberger
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D016875:tau Proteins / Q000138:chemical synthesis* / Q000737:chemistry*
chemical_list: D000345:Affinity Labels; D016875:tau Proteins; D001710:Biotin
keywords: Affinity purification; Expressed protein ligation; Photocleavable biotin; Semi-synthesis; Traceless linker