Inhibition and biochemical studies of cancer-relevant 2-oxoglutarate oxygenase AspH

天冬氨酸/天冬酰胺-β-羟化酶（AspH）是2-酮戊二酸（2 OG）依赖性加氧酶，其催化表皮生长因子样结构域（EGFD）中天冬氨酸和天冬酰胺残基的β-羟基化。AspH在几种人类癌细胞类型中高度过表达，其敲除已被证明可以限制肿瘤生长和转移扩散。人们认为，选择性抑制AspH催化可能是一种新型小分子抗癌治疗剂的成功作用模式。该项目将专注于设计有效的AspH抑制剂，特别是那些针对底物结合口袋的抑制剂，因为这应该会导致相关的2 OG依赖性加氧酶的选择性。另外两个副项目涉及合成一种荧光AspH探针，从而开发出一种基于亲和力的荧光偏振测定法，以及制备含有二硒键的EGFD，这将允许对AspH底物选择性进行生物化学研究。

Aspartate/asparagine-beta-hydroxylase (AspH) is a 2-oxogluturate (2OG)-dependant oxygenase that catalyses the beta-hydroxylation of aspartate and asparagine residues in epidermal growth factor-like domains (EGFDs). AspH is highly overexpressed in several human cancer cell types, and its knockdown has been shown to limit tumour growth and metastatic spread. It is thought selective inhibition of AspH catalysis may be a successful mode of action for a novel, small-molecule anti-cancer therapeutic. This project will focus on the design of efficient AspH inhibitors, in particular those that target the substrate-binding pocket, as this should result in selectivity over related 2OG-dependent oxygenases. Two additional side projects involve the synthesis of a fluorescent AspH probe leading to the development of an affinity-based fluorescence polarization assay, and the preparation of diselenide bond-containing EGFDs that will allow biochemical studies into AspH substrate selectivity.This project falls within the EPSRC "Chemical biology and biological chemistry" research area.