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Updated: Mar 20, 2026

Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange ABE
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Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange ABE

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Control of the signaling of RAS proteins by modulating their palmitoylation

Jiakai Zhu1, Ruiying Guo2, Qi Hu2

  • 1Fudan University, Shanghai 200433, China; Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang 310024, China; State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310024, China; Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang 310024, China.

Cell Chemical Biology
|March 18, 2026
PubMed

Insights

Researchers developed a novel method to control RAS protein palmitoylation using a fusion protein strategy. This approach enables the screening of small molecules that regulate NRAS palmitoylation, offering new tools for cancer research.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • Small GTPases, including RAS proteins, are crucial for cell growth and cancer development.
  • S-palmitoylation, a key post-translational modification, regulates small GTPase localization and function.
  • Targeting S-palmitoylation is challenging due to the complexity of ZDHHC enzymes.

Purpose of the Study:

  • To develop a novel chemical biology tool to modulate RAS protein S-palmitoylation.
  • To establish a high-throughput screening assay for identifying regulators of NRAS palmitoylation.
  • To identify small molecules that inhibit the ZDHHC9-GCP16 complex involved in RAS palmitoylation.

Main Methods:

  • Engineered a fusion protein by linking acyl-protein thioesterase 1 (APT1) to the N-terminus of RAS proteins.
  • Utilized an APT1 inhibitor (ML348) to reversibly control S-palmitoylation and subcellular localization of the fusion protein.
  • Developed and implemented a cell-based high-throughput screening assay to identify NRAS palmitoylation inhibitors.

Main Results:

  • Demonstrated reversible control of RAS palmitoylation and localization via the APT1-RAS fusion protein and ML348.
  • Identified six novel small-molecule compounds that inhibit the ZDHHC9-GCP16 complex.
  • Determined IC50 values for the identified inhibitors, ranging from 1.4 to 8.0 μM.

Conclusions:

  • The APT1-RAS fusion system provides a versatile platform for studying S-palmitoylation.
  • This approach facilitates the discovery of inhibitors targeting the ZDHHC9-GCP16 complex.
  • The findings offer new avenues for therapeutic strategies targeting RAS-driven oncogenesis.
Keywords:
APT1GCP16HRASKRAS4aNRASRASS-acylationZDHHC9depalmitoylationpalmitoylation

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