Efficacy of different NGR tags on Pyrococcus furiosus rubredoxin to bind the CD-13 receptor on mammalian fibrosarcoma cells
Location
Suwanee, GA
Start Date
6-5-2025 1:00 PM
End Date
6-5-2025 4:00 PM
Description
Introduction: In the United States, cancer continues to be a leading cause of death, second to heart disease. The high incidence of death has led to the interest into alternative methods to current-day treatments like chemotherapy, which often have significant side effects and adverse effects on the body. The NGR tag is a peptide sequence closely interacting with APN/CD13 receptors on specific cancer cell lines. Due to this close association, NGR tags have been used to target potential anti-cancer drugs. Pyrococcus furiosus, an anaerobic and hyperthermophilic archaeon, optimally grows in extreme environments. This species has rubredoxin, a small iron-binding protein that is highly stable and can incorporate different metal cofactors.
Objective: The focus of this research project is to further investigate the efficacy and ability of recombinant, Ruthenium-containing Rubredoxin protein containing an NGR tag to bind the CD-13 receptor on HT-1080 Fibrosarcoma cells. The goal is to successfully integrate Ruthenium into the molecular structure of the protein and to treat the Fibrosarcoma cells with varying concentrations of the protein to view the viability and growth of the cells.
Methods: The mutant plasmid with a KNGREG tag at D20 was introduced to Escherichia coli cells via transformation. The protein was expressed in cells grown in a defined media then purified through column chromatography, employing ion exchange and size exclusion to purify the Ru-containing protein. The efficacy of protein purification was tested via SDS-PAGE gel, UV Vis spectroscopy, and mass spectroscopy analysis. This protein will be compared to a version with a CNGRCG tag and to wild-type Rubredoxin to test the ability to bind to APN/CD-13.
Conclusion: Previous work has indicated some positive results of selective binding of the CNGRCG-tagged rubredoxin protein which can be used to investigate targeted chemotherapy. Further research is required to better understand the binding of NGR-APN/CD13 in other mammalian cancer cell lines. Successful binding can lead the way to creating more specific cancer treatments with reduced side effects and lead to a healthier life for cancer patients.
Embargo Period
5-19-2025
Efficacy of different NGR tags on Pyrococcus furiosus rubredoxin to bind the CD-13 receptor on mammalian fibrosarcoma cells
Suwanee, GA
Introduction: In the United States, cancer continues to be a leading cause of death, second to heart disease. The high incidence of death has led to the interest into alternative methods to current-day treatments like chemotherapy, which often have significant side effects and adverse effects on the body. The NGR tag is a peptide sequence closely interacting with APN/CD13 receptors on specific cancer cell lines. Due to this close association, NGR tags have been used to target potential anti-cancer drugs. Pyrococcus furiosus, an anaerobic and hyperthermophilic archaeon, optimally grows in extreme environments. This species has rubredoxin, a small iron-binding protein that is highly stable and can incorporate different metal cofactors.
Objective: The focus of this research project is to further investigate the efficacy and ability of recombinant, Ruthenium-containing Rubredoxin protein containing an NGR tag to bind the CD-13 receptor on HT-1080 Fibrosarcoma cells. The goal is to successfully integrate Ruthenium into the molecular structure of the protein and to treat the Fibrosarcoma cells with varying concentrations of the protein to view the viability and growth of the cells.
Methods: The mutant plasmid with a KNGREG tag at D20 was introduced to Escherichia coli cells via transformation. The protein was expressed in cells grown in a defined media then purified through column chromatography, employing ion exchange and size exclusion to purify the Ru-containing protein. The efficacy of protein purification was tested via SDS-PAGE gel, UV Vis spectroscopy, and mass spectroscopy analysis. This protein will be compared to a version with a CNGRCG tag and to wild-type Rubredoxin to test the ability to bind to APN/CD-13.
Conclusion: Previous work has indicated some positive results of selective binding of the CNGRCG-tagged rubredoxin protein which can be used to investigate targeted chemotherapy. Further research is required to better understand the binding of NGR-APN/CD13 in other mammalian cancer cell lines. Successful binding can lead the way to creating more specific cancer treatments with reduced side effects and lead to a healthier life for cancer patients.