title: Covalent Protein Labeling by Enzymatic Phosphocholination.
abstract: We present a new protein labeling method based on the covalent enzymatic phosphocholination of a specific octapeptide amino acid sequence in intact proteins. The bacterial enzyme AnkX from Legionella pneumophila has been established to transfer functional phosphocholine moieties from synthetically produced CDP-choline derivatives to N-termini, C-termini, and internal loop regions in proteins of interest. Furthermore, the covalent modification can be hydrolytically removed by the action of the Legionella enzyme Lem3. Only a short peptide sequence (eight amino acids) is required for efficient protein labeling and a small linker group (PEG-phosphocholine) is introduced to attach the conjugated cargo.
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department Chemistry, Lichtenbergstrasse 4, 85748 Garching (Germany).
authors.forename: Katharina
authors.initials: K
authors.lastname: Heller
authors.affiliation: Chemical Biology Center (KBC), Institute of Chemistry, Umeå University, 90187 Umeå (Sweden).
authors.forename: Philipp
authors.initials: P
authors.lastname: Ochtrop
authors.affiliation: Chemical Biology Center (KBC), Institute of Chemistry, Umeå University, 90187 Umeå (Sweden).
authors.forename: Michael F
authors.initials: MF
authors.lastname: Albers
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department Chemistry, Lichtenbergstrasse 4, 85748 Garching (Germany).
authors.forename: Florian B
authors.initials: FB
authors.lastname: Zauner
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department Chemistry, Lichtenbergstrasse 4, 85748 Garching (Germany). aymelt.itzen@tum.de.
authors.forename: Aymelt
authors.initials: A
authors.lastname: Itzen
authors.affiliation: Chemical Biology Center (KBC), Institute of Chemistry, Umeå University, 90187 Umeå (Sweden). christian.hedberg@umu.se.|Max Planck Institute of Molecular Physiology, Department of Chemical Biology, Otto-Hahn-Strasse 11, 44227 Dortmund (Germany). christian.hedberg@umu.se.
authors.forename: Christian
authors.initials: C
authors.lastname: Hedberg
mesh_terms: D017089:Ankyrin Repeat; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D006867:Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D016952:Legionella pneumophila / Q000201:enzymology*; D008958:Models, Molecular; D010767:Phosphorylcholine / Q000737:chemistry*; D011499:Protein Processing, Post-Translational
chemical_list: D001426:Bacterial Proteins; D010767:Phosphorylcholine; D006867:Hydrolases
keywords: enzymes; nucleotides; phosphocholination; protein modifications