Enzymatic phosphocholination as a tool for protein labeling
2015 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 282, 12-12 p.Article in journal, Meeting abstract (Other academic) Published
Posttranslational modiﬁcation (PTM) of proteins is a versatile cellular process to regulate the activities of proteins. The high regioselectivity and catalysis rate of posttranslationally modifying enzymes utilizing high-energy precursors can potentially be exploited to equip proteins or peptide sequences with a label of choice site selectively and efﬁciently. We and others have recently described and analyzed a new reversible PTM called phosphocholination in which a phosphocholine group is transferred from a cytidine diphosphate choline (CDP-choline) to a serine residue of the small GTPase Rab1 [1–3]. The enzymes AnkX and Lem3 catalyze the modiﬁcation and the corresponding demodiﬁcation reactions, respectively. Interestingly, we could demonstrate that the modifying enzyme AnkX only requires a short amino acid sequence for substrate recognition. Therefore, we envision AnkX as a tool for the site directed labeling of target proteins. Here we report on the progress of developing a novel reversible protein labeling strategy based on the enzymes AnkX and Lem3 and on derivatives of CDP-choline. We demonstrate the optimization of AnkX and Lem3 enzyme activities and the identiﬁcation of optimal and minimal peptide target sequences. Results indicate that indeed arbitrary proteins of interest can be functionalized with phosphocholine derivatives. In summary, this work yields ﬁrst insights into the development of a CDP-choline based fully reversible protein labeling strategy.
Place, publisher, year, edition, pages
2015. Vol. 282, 12-12 p.
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:umu:diva-111160DOI: 10.1111/febs.13320ISI: 000362570600031OAI: oai:DiVA.org:umu-111160DiVA: diva2:868401
40th Congress of the Federation-of-European-Biochemical-Societies (FEBS) - The Biochemical Basis of Life, Berlin, July 4-9, 2015
Special Issue: SI
Meeting Abstract: P14-004-SH2015-11-102015-11-062015-11-10Bibliographically approved