ArticleNature protocols2014
Single-cell in situ imaging of palmitoylation in fatty-acylated proteins.
Article in Nature protocols, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed.
- Recent advances in S-palmitoylation and its emerging roles in human diseases.Journal of hematology & oncology · 2025Review
- Protocol for the visualization and identification of S-palmitoylated proteins in HCT-116 cells using metabolic labeling via click chemistry.STAR protocols · 2025Article
- ZDHHC2 promoted antimycobacterial responses by selective autophagic degradation of B-RAF and C-RAF in macrophages.Science advances · 2025Article
- Spatial imaging of glycoRNA in single cells with ARPLA.Nature biotechnology · 2024Article
- Advances of Protein Palmitoylation in Tumor Cell Deaths.Cancers · 2023Review
- FAF1 blocks ferroptosis by inhibiting peroxidation of polyunsaturated fatty acids.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Protocol to quantify palmitoylation of cysteines in budding yeast.STAR protocols · 2022Article
- Protein Lipidation Types: Current Strategies for Enrichment and Characterization.International journal of molecular sciences · 2022Review
- Molecular probes for cellular imaging of post-translational proteoforms.RSC chemical biology · 2022Review
- Fatty acylation enhances the cellular internalization and cytosolic distribution of a cystine-knot peptide.iScience · 2021Article
- A small molecule modulating monounsaturated fatty acids and Wnt signaling confers maintenance to induced pluripotent stem cells against endodermal differentiation.Stem cell research & therapy · 2021Article
- A Not-So-Ancient Grease History: Click Chemistry and Protein Lipid Modifications.Chemical reviews · 2021Review
- Determination of the membrane topology of PORCN, an O-acyl transferase that modifies Wnt signalling proteins.Open biology · 2021Article
- Click Chemistry for ImagingBio-protocol · 2021Article
- Regulation of Dynamic Protein S-Acylation.Frontiers in molecular biosciences · 2021Review
- TMEM132A, a Novel Wnt Signaling Pathway Regulator Through Wntless (WLS) Interaction.Frontiers in cell and developmental biology · 2020Article
- Ultrasensitive, multiplexed chemoproteomic profiling with soluble activity-dependent proximity ligation.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- A Hybrid Detection Method Based on Peroxidase-mediated Signal Amplification and Click Chemistry for Highly Sensitive Background-free Immunofluorescent Staining.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2019Article
- Chemoproteomic Method for Profiling Inhibitor-Bound Kinase Complexes.Journal of the American Chemical Society · 2019Article
- Protein Lipidation in Cell Signaling and Diseases: Function, Regulation, and Therapeutic Opportunities.Cell chemical biology · 2018Review
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Dissecting the subcellular distribution of a fatty-acylated protein is key to the understanding of the molecular mechanisms regulating protein movement and function in a cell. This protocol describes how to perform single-cell imaging of palmitoylation in a fatty-acylated protein of interest with high sensitivity using click chemistry, proximity ligation and fluorescence microscopy. The initial steps in this protocol involve optimization of conditions for (i) metabolic incorporation of an alkynyl analog of palmitic acid into cellular proteins coupled with click chemistry and (ii) detecting a specific protein of interest with primary antibodies using automated fluorescence microscopy, followed by (iii) imaging palmitoylation of the target fatty-acylated protein of interest, such as Wnt, Sonic Hedgehog or H-Ras. Furthermore, we outline strategies for imaging specific fatty-acylated proteins with subcellular organelles and/or total proteome palmitoylation, and we discuss special considerations that need to be given depending on the experimental design. The use of clickable fatty acids with proximity ligation may have promising applications to the investigation of fatty acylation cell biology. The entire protocol takes ∼3 weeks to complete.
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