Nudix Hydrolases and HAD Superfamily Aromatic Phosphatases

Nudix 水解酶和 HAD 超家族芳香族磷酸酶

• 批准号: 7127830
• 负责人: SUZANNE Freeman O'HANDLEY
• 金额: $ 20.72万
• 依托单位: ROCHESTER INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-17 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7127830
• 关键词: Mycobacterium leprae; Mycobacterium tuberculosis; acid phosphatase; bacterial cytopathogenic effect; bacterial genetics; enzyme activity; enzyme mechanism; expression cloning; nucleotidases; operon; protein purification; site directed mutagenesis

DESCRIPTION (provided by applicant): The specific aims of this research are to discover and characterize new enzymes of the Nudix hydrolase superfamily and the aromatic phosphatase family of the HAD (Haloacid Dehalogenase) superfamily. The Nudix hydrolases are defined as enzymes that hydrolyze substrates consisting of a nucleoside diphosphate linked to some moiety x, and contain the signature sequence GX5EX7REUXEEXGU (U=l, L, or V). Nudix substrates include nucleoside triphosphates, nucleotide coenzymes, nucleotide sugars, diadenosine polyphosphates, and CDP-alcohols, and the Nudix hydrolases appear to perform vital cellular functions including removal of toxic compounds and modulation of natural metabolites to regulate signaling or biochemical pathways. We will focus on the Nudix hydrolases from Mycobacterium tuberculosis and Mycobacterium leprae, as potential novel antibiotic targets. The HAD superfamily is diverse, consisting of haloacid dehalogenases, phosphonatases, phosphomutases, and a number of distinctive phosphatase families. We will focus on a little defined phosphatase family within this superfamily, the aromatic phosphatases, formerly known as the nitrophenyl phosphatase family. Because only five enzymes with three activities have been identified for this family to date, the opportunity for discovery of new enzymatic activities is high. Concentrating on enzymes within operons or with known phenotypes will increase our success rate. Members of these two families will be cloned, expressed, purified, and characterized. Characterization will include biochemical analysis to determine substrate specificity and enzymatic activity, knockout mutagenesis with phenotypic and complementation studies to identify cellular function, and structural determination to understand structure-function relationships and for future drug design. Many of the enzymes discovered thus far appear to have important biological functions, thus making them good candidates as potential novel antibiotic targets in pathogenic bacteria.

描述（由申请人提供）：本研究的具体目的是发现和表征 Nudix 水解酶超家族的新酶和 HAD（卤酸脱卤酶）超家族的芳香族磷酸酶家族。 Nudix水解酶被定义为水解由与某些x部分连接的核苷二磷酸组成的底物的酶，并且包含特征序列GX5EX7REUXEEXGU(U＝1、L或V)。 Nudix 底物包括三磷酸核苷、核苷酸辅酶、核苷酸糖、多磷酸二腺苷和 CDP 醇，Nudix 水解酶似乎执行重要的细胞功能，包括去除有毒化合物和调节天然代谢物以调节信号传导或生化途径。我们将重点关注来自结核分枝杆菌和麻风分枝杆菌的 Nudix 水解酶，作为潜在的新型抗生素靶点。 HAD 超家族多种多样，由卤酸脱卤酶、磷酸酶、磷酸变位酶和许多独特的磷酸酶家族组成。我们将重点关注这个超家族中一个稍微明确的磷酸酶家族，即芳香族磷酸酶，以前称为硝基苯基磷酸酶家族。由于迄今为止该家族仅鉴定出五种具有三种活性的酶，因此发现新酶活性的机会很高。专注于操纵子内的酶或具有已知表型的酶将提高我们的成功率。这两个家族的成员将被克隆、表达、纯化和表征。表征将包括生化分析以确定底物特异性和酶活性，通过表型和互补研究进行敲除突变以识别细胞功能，以及结构测定以了解结构-功能关系并用于未来的药物设计。迄今为止发现的许多酶似乎具有重要的生物学功能，因此使它们成为病原菌中潜在的新型抗生素靶点的良好候选者。

项目成果

YZGD from Paenibacillus thiaminolyticus, a pyridoxal phosphatase of the HAD (haloacid dehalogenase) superfamily and a versatile member of the Nudix (nucleoside diphosphate x) hydrolase superfamily.

YZGD 来自类硫胺素类芽孢杆菌，HAD（卤酸脱卤酶）超家族的吡哆醛磷酸酶，也是 Nudix（核苷二磷酸 x）水解酶超家族的多功能成员。

• DOI: 10.1042/bj20051172
• 发表时间: 2006
• 期刊: The Biochemical journal
• 作者: Tirrell,IsaacM;Wall,JenniferL;Daley,ChristopherJ;Denial,SarahJ;Tennis,FrancesG;Galens,KevinG;O'Handley,SuzanneF
• 通讯作者: O'Handley,SuzanneF