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Tuning PFKFB3 Bisphosphatase Activity Through Allosteric Interference.
Helena Macut1, Xiao Hu1, Delia Tarantino2
1DISFARM- Department of Pharmaceutical sciences, Via Mangiagalli 25, 20133, Milan, Italy.
Scientific Reports
|January 1, 2020
Summary
Researchers discovered a novel method to modulate phosphofructokinase-2,6-bisphosphatase (PFKFB3) activity by activating its allosteric bisphosphatase site with small peptides, offering a new metabolic regulation strategy.
Area of Science:
- Biochemistry
- Enzymology
- Metabolic Regulation
Background:
- PFKFB3 is a key regulator of cellular metabolism, controlling fructose-2,6-bisphosphate levels.
- It possesses a high kinase-to-phosphatase activity ratio, making direct kinase inhibition a common strategy.
- An indirect modulation approach targeting the bisphosphatase activity is explored.
Purpose of the Study:
- To develop an innovative strategy for modulating PFKFB3 activity via allosteric bisphosphatase activation.
- To discover and synthesize small peptides targeting an allosteric site on PFKFB3.
- To establish a method for assessing PFKFB3 bisphosphatase activity and peptide-induced activation.
Main Methods:
- Peptide library synthesis and screening.
- Microscale Thermophoresis (MST) for binding affinity evaluation.
- LC-MS/MS method development for bisphosphatase activity assay.
- Experimental and computational investigations of allosteric site-activity relationship.
Main Results:
- Discovery and synthesis of PFKFB3-binding peptides targeting an allosteric site.
- Successful development of an LC-MS/MS method to quantify PFKFB3 bisphosphatase activity.
- Demonstration of peptide-induced activation of PFKFB3 bisphosphatase activity.
- Elucidation of the molecular mechanism linking the allosteric site to bisphosphatase function.
Conclusions:
- A novel indirect strategy for PFKFB3 modulation through allosteric bisphosphatase activation has been established.
- Small peptides can effectively target an allosteric site to modulate PFKFB3 activity.
- The developed LC-MS/MS method provides a robust tool for assessing PFKFB3 bisphosphatase activity and modulation.
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