Dipartimento di Biologia, Università Federico II, 80126 Napoli, Italy.
Dipartimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Università degli Studi della Campania "Luigi Vanvitelli", 81100 Caserta, Italy.
Int J Mol Sci. 2019 Aug 26;20(17):4164. doi: 10.3390/ijms20174164.
A large number of mutations causing PMM2-CDG, which is the most frequent disorder of glycosylation, destabilize phosphomannomutase2. We looked for a pharmacological chaperone to cure PMM2-CDG, starting from the structure of a natural ligand of phosphomannomutase2, α-glucose-1,6-bisphosphate. The compound, β-glucose-1,6-bisphosphate, was synthesized and characterized via P-NMR. β-glucose-1,6-bisphosphate binds its target enzyme in silico. The binding induces a large conformational change that was predicted by the program PELE and validated in vitro by limited proteolysis. The ability of the compound to stabilize wild type phosphomannomutase2, as well as frequently encountered pathogenic mutants, was measured using thermal shift assay. β-glucose-1,6-bisphosphate is relatively resistant to the enzyme that specifically hydrolyses natural esose-bisphosphates.
大量导致 PMM2-CDG 的突变使磷酸甘露糖变位酶 2 不稳定，PMM2-CDG 是最常见的糖基化紊乱。我们从磷酸甘露糖变位酶 2 的天然配体α-葡萄糖-1,6-双磷酸的结构出发，寻找一种治疗 PMM2-CDG 的药理学伴侣。通过 P-NMR 合成并表征了化合物β-葡萄糖-1,6-双磷酸。该化合物在计算机中与靶酶结合。该结合诱导了一个大的构象变化，该变化由程序 PELE 预测，并通过有限的蛋白水解在体外验证。通过热移位测定测量了该化合物稳定野生型磷酸甘露糖变位酶 2 以及常见致病性突变体的能力。β-葡萄糖-1,6-双磷酸对专门水解天然己糖-双磷酸的酶具有相对抗性。
