title: Traceless purification and desulfurization of tau protein ligation products.
abstract: We present a novel strategy for the traceless purification and synthetic modification of peptides and proteins obtained by native chemical ligation. The strategy involves immobilization of a photocleavable semisynthetic biotin-protein conjugate on streptavidin-coated agarose beads, which eliminates the need for tedious rebuffering steps and allows the rapid removal of excess peptides and additives. On-bead desulfurization is followed by delivery of the final tag-free protein product. The strategy is demonstrated in the isolation of a tag-free Alzheimer's disease related human tau protein from a complex EPL mixture as well as a triphosphorylated peptide derived from the C-terminus of tau.
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany); Humboldt Universität zu Berlin, Department Chemie, Brook-Taylor-Strasse 2, 12489 Berlin (Germany); Freie Universität Berlin, Institut für Chemie und Biochemie, Takustrasse 3, 14195 Berlin (Germany).
authors.forename: Oliver
authors.initials: O
authors.lastname: Reimann
authors.forename: Caroline
authors.initials: C
authors.lastname: Smet-Nocca
authors.forename: Christian P R
authors.initials: CP
authors.lastname: Hackenberger
mesh_terms: D000595:Amino Acid Sequence; D001710:Biotin / Q000737:chemistry; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010455:Peptides; D013455:Sulfur / Q000302:isolation & purification*; D014466:Ultraviolet Rays; D016875:tau Proteins / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D055767:Immobilized Proteins; D010455:Peptides; D016875:tau Proteins; D001710:Biotin; D013455:Sulfur
keywords: desulfurization; native chemical ligation; peptide purification; photocleavable biotin; traceless linkers