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Updated: Jun 13, 2026
Visualization of Protein-protein Interaction in Nuclear and Cytoplasmic Fractions by Co-immunoprecipitation and In Situ Proximity Ligation Assay
Published on: January 16, 2017
Proteomic analysis reveals novel binding partners of MIP-T3 in human cells
Chao-Wan Guo1, Sheng Xiong, Ge Liu
1Division of Molecular Pharmacology of Infectious agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki, Japan.
Abstract:
MIP-T3 (microtubule-interacting protein associated with TRAF3) is a microtubule-interacting protein that evolutionarily conserved from worms to humans, but whose cellular functions remains unknown. To get insight into the functions of MIP-T3, we set out to identify MIP-T3 interacting proteins by immunoprecipitation in human embryonic kidney 293 cells and MS analysis. As the results, a total of 34 proteins were identified and most of them were novel MIP-T3 putative partners. The MIP-T3-associated proteins could be grouped into nine clusters based on their molecule functions, including cytoskeleton, chaperone, nucleic acid binding, kinase and so on. Three MIP-T3-interacted proteins - actin, HSPA8 and tubulin - were further confirmed by reciprocal coimmunoprecipitations and colocalization analysis. The interaction of MIP-T3 with both actin filaments and microtubule suggested that MIP-T3 may play an important role in regulation of cytoskeleton dynamics in cells. Our results therefore not only uncover a large number of MIP-T3-associated proteins that possess a variety of cellular functions, but also provide new research directions for the study of the functions of MIP-T3.
Insights
Microtubule-interacting protein associated with TRAF3 (MIP-T3) interacts with cytoskeleton proteins like actin and tubulin. This discovery offers new insights into MIP-T3
Area of Science:
- Cell Biology
- Molecular Biology
- Protein Interactions
Background:
- Microtubule-interacting protein associated with TRAF3 (MIP-T3) is an evolutionarily conserved protein.
- The specific cellular functions of MIP-T3 remain largely unknown.
Purpose of the Study:
- To identify proteins that interact with MIP-T3.
- To gain insights into the cellular functions of MIP-T3.
Main Methods:
- Immunoprecipitation in human embryonic kidney 293 cells.
- Mass spectrometry (MS) analysis.
- Reciprocal co-immunoprecipitations and colocalization analysis for validation.
Main Results:
- Identified 34 novel putative MIP-T3 interacting partners.
- MIP-T3-associated proteins involved in cytoskeleton, chaperone, nucleic acid binding, and kinase functions.
- Confirmed interactions with actin, HSPA8, and tubulin, suggesting a role in cytoskeleton dynamics.
Conclusions:
- Uncovered a broad range of MIP-T3-associated proteins.
- MIP-T3 likely plays a significant role in regulating cytoskeleton dynamics.
- Provides new research avenues for understanding MIP-T3 functions.
