Identification and characterization of tubulin as Ga(III)-binding protein in T24 cells
Junxin Chen1, Xue Yang1, Shuaile Jia1
1Hebei Key Laboratory for Chronic Diseases, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan, Hebei 063210, China.
Abstract:
Gallium-based metallic drugs and agents have been widely applied for the diagnosis and treatment of diseases such as non-Hodgkin's lymphoma (NHL), but there are few reports on the potential Ga(III)-binding proteins and the related cytotoxic mechanisms for Ga(III). Herein, by using human urinary bladder cancer T24 cells as a model, we identify and report that tubulin is a Ga(III)-binding protein target in T24 cells. Our analyses, including the employment of a series of methods based on immobilized metal affinity chromatography (IMAC), cellular thermal shift assay (CETSA), and immunofluorescence experiments, collectively explained this finding. Our results suggest that the binding of Ga(III) to tubulin led to significant changes in the morphology and distribution of microtubules in cells. The blocked microtubule formation or microtubule depolymerization as a result of the binding of Ga(III) to tubulin may be an important molecular mechanism by which Ga(III) exerts its cytotoxic effects in T24 cells to inhibit tumor cell growth.
Insights
Gallium(III) (Ga(III)) binds to tubulin in bladder cancer cells, disrupting microtubule formation. This interaction is a key mechanism for Ga(III) anticancer effects, inhibiting tumor growth.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Gallium compounds are used in cancer diagnosis and treatment, notably for non-Hodgkin's lymphoma (NHL).
- Understanding gallium's molecular targets and cytotoxic mechanisms is crucial for developing effective cancer therapies.
- Limited information exists regarding specific gallium(III) (Ga(III))-binding proteins and their roles in cancer cell death.
Purpose of the Study:
- To identify Ga(III)-binding proteins in human urinary bladder cancer T24 cells.
- To elucidate the cytotoxic mechanisms of Ga(III) in bladder cancer.
- To investigate the interaction between Ga(III) and its protein targets at the cellular level.
Main Methods:
- Immobilized metal affinity chromatography (IMAC) for protein binding analysis.
- Cellular thermal shift assay (CETSA) to confirm target engagement.
- Immunofluorescence microscopy to visualize cellular and subcellular changes.
- Analysis of microtubule morphology and distribution.
Main Results:
- Tubulin was identified as a direct Ga(III)-binding protein target in T24 bladder cancer cells.
- Ga(III) binding to tubulin induced significant alterations in microtubule structure and organization.
- Microtubule formation was inhibited, and depolymerization was observed upon Ga(III) exposure.
Conclusions:
- Tubulin is a critical molecular target for Ga(III) in bladder cancer cells.
- Disruption of microtubule dynamics by Ga(III) binding to tubulin is a primary mechanism of its cytotoxic action.
- These findings provide insights into Ga(III)-based cancer therapy and highlight tubulin as a target for inhibiting tumor cell proliferation.
More Related Videos
06:30Author Spotlight: Purifying High-Quality Tubulin to Study Protein Dynamics and Therapeutic Applications
Published on: October 11, 2024
07:21Quantitative Microtubule Fractionation Technique to Separate Stable Microtubules, Labile Microtubules, and Free Tubulin in Mouse Tissues
Published on: November 17, 2023