Covalent Protein Labeling by Enzymatic Phosphocholination |
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| Authors: | MSc Katharina Heller MSc Philipp Ochtrop MSc Michael F Albers Florian B Zauner Prof?Dr Aymelt Itzen Prof?Dr Christian Hedberg |
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| Affiliation: | 1. Center for Integrated Protein Science Munich, Technische Universit?t München, Department Chemistry, Lichtenbergstrasse 4, 85748 Garching (Germany);2. Chemical Biology Center (KBC), Institute of Chemistry, Ume? University, 90187 Ume? (Sweden);3. Max Planck Institute of Molecular Physiology, Department of Chemical Biology, Otto‐Hahn‐Strasse 11, 44227 Dortmund (Germany) |
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| 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. |
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| Keywords: | enzymes nucleotides phosphocholination protein modifications |
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