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Zonula occludin toxin, a microtubule binding protein
World Journal of Gastroenterology
|January 31, 2002
Summary
Zot protein interacts with intracellular microtubules (Mt) to regulate intestinal barrier permeability. This binding activates signaling pathways, leading to the opening of intercellular tight junctions.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Zot is a protein involved in regulating intestinal permeability.
- Microtubules (Mt) play crucial roles in maintaining cell structure and function.
- The precise mechanism by which Zot influences intestinal tight junctions remains incompletely understood.
Purpose of the Study:
- To investigate the interaction between Zot and microtubules.
- To identify Zot-binding proteins within intestinal cells.
- To elucidate the role of Zot-microtubule interaction in regulating intestinal tight junction permeability.
Main Methods:
- Zot affinity chromatography was used to purify Zot-binding proteins from intestinal cell lysates.
- N-terminal sequencing was performed on eluted protein bands.
- Western blotting with anti-Zot and anti-tau antibodies was employed.
- In vitro tubulin binding assays were conducted.
Main Results:
- Two protein bands (60 kDa and 45 kDa) were identified as Zot-binding proteins.
- The 60 kDa band's N-terminal sequence matched beta-tubulin.
- Zot showed cross-reactivity with anti-tau antibodies.
- Zot co-precipitated with microtubules in vitro, confirming tubulin-binding properties.
Conclusions:
- Zot binds to intracellular microtubules (Mt).
- This interaction is proposed to regulate intestinal tight junction permeability.
- Zot binding to Mt likely activates intracellular signaling pathways, leading to tight junction permeabilization.