Related Experiment Video
Updated: Jun 10, 2026
04:56
Detection of Aggregation-Prone Behavior in Mutant P53 V157F Breast Cancer Cells Using Multipoint Thioflavin T Fluorescence
Published on: December 30, 2025
Isolation of a point-mutated p47 lacking binding affinity to p97ATPase
Yayoi Kaneko1, Kaori Tamura, Go Totsukawa
1Department of Molecular Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
FEBS Letters
|August 10, 2010
Summary
Researchers created a mutated p47 protein, F253S, that cannot bind to p97. This mutation disrupted Golgi membrane fusion and cisternal regrowth, highlighting p47
Area of Science:
- Cell Biology
- Molecular Biology
- Protein Interactions
Background:
- p47 is a p97-binding protein crucial for Golgi membrane fusion.
- It collaborates with p97 and VCIP135 in this process.
Purpose of the Study:
- To investigate the role of p97-binding affinity in p47's function.
- To elucidate the specific p97-binding regions of p47.
Main Methods:
- Site-directed mutagenesis to create p47 F253S mutant.
- Co-immunoprecipitation to assess p97-p47 complex formation.
- In vitro Golgi reassembly assay to evaluate cisternal regrowth.
Main Results:
- The F253S mutation in p47 abolished its binding affinity to p97.
- p47(F253S) failed to form a complex with p97 and did not induce cisternal regrowth.
- Similar mutations in p37 and Ufd1 also impaired their p97-binding ability.
Conclusions:
- p97-binding is essential for p47's role in Golgi membrane fusion and reassembly.
- The F253S mutation identifies a critical p97-binding site in p47.
- This finding has implications for understanding the broader p97-dependent protein complex functions.
