Monitoring wnt protein acylation using an in vitro cyclo- addition reaction

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Abstract

We describe here a technique for visualizing the lipidation status of Wnt proteins using azide-alkyne cycloaddition chemistry (click chemistry) and SDS-PAGE. This protocol incorporates in vivo labeling of a Wnt- IgG Fc fusion protein using an alkynylated palmitate probe but departs from a traditional approach by incorporating a secondary cycloaddition reaction performed on single-step purified Wnt protein immobilized on protein A resin. This approach mitigates experimental noise by decreasing the contribution of labeling from other palmitoylated proteins and by providing a robust method for normalizing labeling efficiency based on protein abundance.

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Tuladhar, R., Yarravarapu, N., & Lum, L. (2016). Monitoring wnt protein acylation using an in vitro cyclo- addition reaction. In Methods in Molecular Biology (Vol. 1481, pp. 11–16). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6393-5_2

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