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Updated: Jun 9, 2026
14:47
Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry
Published on: May 17, 2016
Mapping the Interactome of c-Myc/Max Heterodimer by Micro-affinity Purification Coupling Mass Spectrometry.
Ying Zhu1,2, Ying Han3, Yue Zhang2,4
1School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
Journal of Proteome Research
|June 8, 2026
Summary
We developed a novel micro-immobilized metal affinity chromatography (m-IMAC) method to map protein-protein interactions (PPIs). This technique identified 574 c-Myc/Max interacting proteins, advancing our understanding of transcriptional regulation and tumorigenesis.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- c-Myc/Max is a critical transcriptional regulator involved in gene expression and tumorigenesis.
- Understanding its protein-protein interactions (PPIs) is crucial for deciphering its regulatory mechanisms.
- Existing methods for interactome mapping can be limited in scope and efficiency.
Purpose of the Study:
- To develop and validate a micro-immobilized metal affinity chromatography (m-IMAC) coupled with label-free quantification (LFQ) method for PPI mapping.
- To comprehensively map the interactome of the c-Myc/Max complex.
- To gain insights into the biological functions associated with c-Myc/Max interactions.
Main Methods:
- Fabrication of m-IMAC columns using 2-hydroxyethyl methacrylate phosphate (HEMAP) and pentaerythritol triacrylate.
- Immobilization of recombinant His6-tagged bait proteins (c-Myc/Max) onto Ni2+-chelating columns.
- Enrichment of interacting proteins from cell lysate followed by data-independent acquisition (DIA)-based LFQ.
- Gene Ontology (GO) analysis of identified proteins.
Main Results:
- Successfully developed and applied the m-IMAC-LFQ method for PPI analysis.
- Identified 574 c-Myc/Max-interacting proteins, with a significant number being novel.
- GO analysis revealed enrichment in RNA processing, chromatin organization, and ribosome biogenesis pathways.
Conclusions:
- The m-IMAC-LFQ method provides a versatile and efficient strategy for high-throughput interactome analysis.
- This study presents the most comprehensive interactome map of c-Myc/Max to date.
- The findings offer new insights into c-Myc/Max-mediated transcriptional regulation and its role in cancer.
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