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Analysis of the Interaction of UBE2Q1 with B4GALT1 and P53: Experimental and Molecular Modeling Study
Hadi Ghasemi1, Atefeh Seghatoleslam1,2, Mohammad Ali Fahmideh Kar3
1Autophagy Research Center, Department of Clinical Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Background:
UBE2Q1-dependent ubiquitination of key proteins including β 1,4- galactosyltransferase (GalT1), and P53 might play a pivotal role in cancer development.
Objective:
The present study aimed to evaluate the molecular analysis of possible interactions between UBE2Q1 with B4GALT1 and P53 proteins.
Methods:
We established SW1116 colorectal cancer cell line stably transfected with UBE2Q1. To verify the overexpression of UBE2Q1, we performed western blot and fluorescent microscopy analysis. Using the immunoprecipitation (IP) product of the over-expressed protein on the silver staining gel, we observed the potential interacting partners of UBE2Q1. The Molecular Operating Environment (MOE) software was also used to perform the molecular docking of the UBC domain of UBE2Q1 (2QGX) with B4GALT1 (2AGD), and P53 (tetramerization (1AIE) and DNA binding domains (1GZH)) proteins.
Results:
Western blot and IP analysis detected a UBE2Q1-GFP band in transfected cells, while no band was detected for mock-transfected cells. Moreover, the overexpression of UBE2Q1 tagged with GFP was observed under fluorescent microscopy as well with about 60-70% shining. Silver staining of IP gel revealed several bands in colorectal cancer (CRC) with UBE2Q1 overexpression. Protein- Protein interaction (PPI) analysis also depicted a high affinity of the UBC domain of UBE2Q1 to the B4GALT1 and P53 (tetramerization and DNA binding domains). Molecular docking also revealed hot-spot regions for all poses.
Conclusion:
Our data suggest that UBE2Q1 as an E2 enzyme of ubiquitination system can interact with B4GALT1 and P53, and may contribute to the accumulation of misfolded important proteins and colorectal tumor development.
Insights
UBE2Q1, an E2 ubiquitination enzyme, interacts with B4GALT1 and P53 proteins. This interaction may contribute to colorectal tumor development by accumulating misfolded proteins.
Area of Science:
- Molecular Biology
- Biochemistry
- Oncology
Background:
- UBE2Q1-dependent ubiquitination of key proteins like β-1,4-galactosyltransferase (GalT1) and P53 is implicated in cancer development.
- Understanding these interactions is crucial for cancer research.
Purpose of the Study:
- To investigate the molecular interactions between UBE2Q1 and B4GALT1 and P53 proteins.
- To analyze the role of UBE2Q1 in colorectal cancer.
Main Methods:
- Established SW1116 colorectal cancer cell line with stable UBE2Q1 overexpression.
- Utilized western blot, fluorescent microscopy, and immunoprecipitation (IP) for protein analysis.
- Performed molecular docking using MOE software to analyze UBE2Q1 (UBC domain) with B4GALT1 and P53.
Main Results:
- Confirmed UBE2Q1 overexpression in transfected cells via western blot and microscopy.
- IP and silver staining identified potential UBE2Q1 interacting partners.
- Molecular docking revealed high affinity and hot-spot regions for UBE2Q1 (UBC domain) binding to B4GALT1 and P53.
Conclusions:
- UBE2Q1, an E2 ubiquitination enzyme, interacts with B4GALT1 and P53.
- These interactions may drive colorectal tumor development through the accumulation of misfolded proteins.
