Related Experiment Video

Updated: Nov 2, 2025

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification BiCAP
06:45

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification BiCAP

Published on: June 15, 2018

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Stable and EGF-Induced Temporal Interactome Profiling of CBL and CBLB Highlights Their Signaling Complex Diversity

Jie Liu1,2, Lijun Yang1,2, An He3

  • 1Department of Oncology, The Second Clinical Medical College, Jinan University (Shenzhen People's Hospital), Shenzhen 518020, China.

Journal of Proteome Research
|June 17, 2021
PubMed

Insights

This study identifies proteins interacting with CBL and CBLB, crucial regulators of epidermal growth factor receptor (EGFR) signaling. The findings reveal redundant and complementary functions of CBL and CBLB in controlling cancer-related cell activities.

Area of Science:

  • Cell Biology
  • Molecular Oncology
  • Signal Transduction

Background:

  • Epidermal growth factor receptor (EGFR) signaling is vital for cell functions but its aberrant activation drives cancer.
  • CBL proteins (CBL and CBLB) negatively regulate EGFR signaling through ubiquitination and degradation, controlling signal duration and intensity.

Purpose of the Study:

  • To identify and characterize the protein interactomes of CBL and CBLB.
  • To understand the dynamic assembly of signaling proteins in response to EGFR activation.
  • To elucidate the functional relationship between CBL and CBLB in EGFR signaling.

Main Methods:

  • Stable inducible expression of FLAG-tagged CBL and CBLB in cell lines.
  • Affinity purification followed by mass spectrometry (AP-MS) to identify interacting proteins.
  • Time-resolved proteomic profiling across six time points.
  • Biochemical assays for validating protein associations.

Main Results:

  • Identified 102 stable interacting proteins for CBL and 82 for CBLB.
  • Temporal profiling revealed dynamic protein assembly during EGFR signaling.
  • Demonstrated redundant and complementary functions of CBL and CBLB.
  • Validated interactions with EPS15L1, ITSN2, and TNK2.

Conclusions:

  • The study provides a comprehensive resource of CBL and CBLB interactomes.
  • Highlights the significant and varied roles of CBL and CBLB in the EGFR signaling network.
  • Offers insights into the regulation of cell proliferation, migration, and survival in cancer contexts.
Keywords:
AP-MSCBLCBLBdynamic protein complexessignal transduction

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