Abstract
Cholesterol modification (or cholesterylation) is a rare but important posttranslational lipid modification of proteins. So far, two proteins, namely Hedgehog and Smoothened, have been characterized to undergo cholesterylation. It is unknown whether other proteins are similarly modified. Here we present a protocol of a chemical biology method to analyze the Smoothened protein cholesterylation using an azido-conjugated cholesterol analog combined with click chemistry. This method can be applied to analyze Hedgehog cholesterylation, and be extended to identify novel cholesterylated proteins with minor changes.
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Acknowledgments
We thank Dr. Jie Luo for critical reading of the manuscript. This work was supported by grants from the National Natural Science Foundation (China; Nos. 91753204 and 31771568) and Ministry of Science and Technology (China; No. 2016YFA0500100). B.-L. Song acknowledges the support from the Tencent Foundation through the XPLORER PRIZE.
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Hu, A., Zhou, M., Song, BL. (2022). Analysis of Protein Cholesterylation by Biorthogonal Labeling. In: Li, X. (eds) Hedgehog Signaling. Methods in Molecular Biology, vol 2374. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1701-4_3
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DOI: https://doi.org/10.1007/978-1-0716-1701-4_3
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