Thermodynamic and structural study on the interaction of an enzyme and a substrate analogue for development of new therapy for lysosomal diseases

为了获得用于Fabry病和Pompe病的伴侣治疗的基本信息，我们采用等温滴定量热法和表面等离子体共振生物传感器检测了α-半乳糖苷酶A /酸性α-葡萄糖苷酶与其底物类似物之间的分子相互作用，并首先确定了热力学和结合动力学参数。此外，硅结构分析表明，这些化学物质适合于酶分子的活性位点口袋，并与构成口袋的残基发生氢键。化学物质的侧链被认为是朝向口袋入口的。在此基础上，我们设计了一种新的化学物质，可以强烈稳定一种突变酶。热力学和结构方面的研究将为改进Fabry病和Pompe病的伴侣治疗提供许多信息。

To obtain basic information useful for development of chaperone therapy for Fabry disease and Pompe disease, we examined the molecular interactions between α-galactosidase A /acid α-glucosidase and their substrate analogues by means of isothermal titration calorimetry and surface plasmon resonance biosensor assays, and first determined the thermodynamic and binding-kinetics parameters. Furthermore, the structural analysis in silico revealed that the chemicals fit into the active site pocket of the enzyme molecule and undergo hydrogen bonding with residues comprising the pocket. The side chain of the chemicals is thought to be oriented towards the entrance of the pocket. On the basis of the results, we designed a new chemical that strongly stabilizes a mutant enzyme. Thermodynamic and structural studies should provide us with a lot of information for improving chaperone therapy for Fabry disease and Pompe disease.