1. Exploring ligand interactions with human phosphomannomutases using recombinant bacterial thermal shift assay and biochemical validation.
- Author
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Monticelli, Maria, Hay Mele, Bruno, Wright, Demi Marie, Guerriero, Simone, Andreotti, Giuseppina, and Cubellis, Maria Vittoria
- Subjects
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MOLECULAR docking , *MOLECULAR chaperones , *SOCIAL interaction , *PHOSPHATASE inhibitors , *CONGENITAL disorders - Abstract
PMM2-CDG, a disease caused by mutations in phosphomannomutase-2, is the most common congenital disorder of glycosylation. Yet, it still lacks a cure. Targeting phosphomannomutase-2 with pharmacological chaperones or inhibiting the phosphatase activity of phosphomannomutase-1 to enhance intracellular glucose-1,6-bisphosphate have been proposed as therapeutical approaches. We used Recombinant Bacterial Thermal Shift Assay to assess the binding of a substrate analog to phosphomannomutase-2 and the specific binding to phosphomannomutase-1 of an FDA-approved drug - clodronate. We also deepened the clodronate binding by enzyme activity assays and in silico docking. Our results confirmed the selective binding of clodronate to phosphomannomutase-1 and shed light on such binding. • PMM2-CDG is a congenital disorder of glycosylation caused by mutations in phosphomannomutase-2 (PMM2). It has no cure. • Alpha- glucose-1,6-bisphosphate is a PMM2 stabilizer and activator, hydrolysed by a phosphatase in vivo. • We evaluated two approaches to develop therapies: pharmacological chaperones to stabilize PMM2 or specific phosphatase inhibitors. • We deepened the effect of Clodronate. Clodronate is a selective inhibitor for the main phosphatase acting on glucose-1,6-bisphosphate. • Recombinant Bacterial Cellular Thermal Shift Assay can be helpful in fast and preliminary drug screening. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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