Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo
Abstract
1. Introduction
2. Materials and Methods
2.1. Generation and Characterization of GRAF3RQ SMMHC-CreERT2 Mice
2.2. Blood Pressure Measurements
2.3. Cell Culture
2.4. Molecular Modeling
2.5. FRET Conformation Assay
2.6. Western Blotting
2.7. Immunoprecipitation
2.8. Protein Purification
2.9. Radioactive In Vitro Kinase assay
2.10. Time Resolved-Fluorescence Energy Transfer (TR-FRET) Assay
2.11. Bioluminescence Resonance Energy Transfer (BRET) Assay
2.12. Statistics
3. Results
3.1. Increased SMC GRAF3 Expression Reduced BP in Mice
3.2. GRAF3 Activity is Dynamically Regulated by Auto-Inhibitory Interactions
3.3. FAK and Src-Mediated Phosphorylation of GRAF3 at Y376 Promotes Allosteric Activation
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Dee, R.A.; Bai, X.; Mack, C.P.; Taylor, J.M. Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo. Cells 2020, 9, 1042. https://doi.org/10.3390/cells9041042
Dee RA, Bai X, Mack CP, Taylor JM. Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo. Cells. 2020; 9(4):1042. https://doi.org/10.3390/cells9041042
Chicago/Turabian StyleDee, Rachel A., Xue Bai, Christopher P. Mack, and Joan M. Taylor. 2020. "Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo" Cells 9, no. 4: 1042. https://doi.org/10.3390/cells9041042
APA StyleDee, R. A., Bai, X., Mack, C. P., & Taylor, J. M. (2020). Molecular Regulation of the RhoGAP GRAF3 and Its Capacity to Limit Blood Pressure In Vivo. Cells, 9(4), 1042. https://doi.org/10.3390/cells9041042