TYPE II NADH: QUINONE OXIDOREDUCTASES OF PLASMODIUM FALCIPARUM AND MYCOBACTERIUM TUBERCULOSIS: KINETIC AND HIGH-THROUGHPUT ASSAYS

被引：33

作者：

Fisher, Nicholas ^{[1
]}

Warman, Ashley J. ^{[1
]}

Ward, Stephen A. ^{[1
]}

Biagini, Giancarlo A. ^{[1
]}

机构：

[1] Univ Liverpool, Liverpool Sch Trop Med, Liverpool L3 5QA, Merseyside, England

来源：

METHODS IN ENZYMOLOGY, VOL 456: MITOCHONDRIAL FUNCTION, PART A: MITOCHONDRIAL ELECTRON TRANSPORT COMPLEXES AND REACTIVE OXYGEN SPECIES | 2009年 / 456卷

基金：

英国惠康基金;

关键词：

MENAQUINONE OXIDOREDUCTASE; ESCHERICHIA-COLI; UBIQUINONE OXIDOREDUCTASE; SCREENING ASSAYS; COMPLEX-I; INHIBITORS; DEHYDROGENASE; MITOCHONDRIA; SEQUENCE; CULTURE;

D O I：

10.1016/S0076-6879(08)04417-0

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Type II NADH: quinone oxidoreductases (ndh) are flavoenzymes found in a broad range of organisms including plants, fungi, protozoa, and bacteria. The ndh enzymes catalyze the oxidation of NADH with concomitant reduction of quinone (Q). These membrane-bound respiratory enzymes differ from the canonical NADH: dehydrogenase (complex 1), because they are not involved in the vectorial transfer of protons across membranes. In Plasmodium falciparum and Mycobacterium tuberculosis, causative agents of malaria and tuberculosis, respectively, ndhs have aroused interest because of the essential role played in maintaining a reduced Q-pool during infection. In this chapter, we present methods for the measurement of steady-state parameters for ndhs from both pathogens, highlighting best practices and caveats. In addition, owing to the interest in ndhs as potential chemotherapeutic targets, we describe a miniaturized endpoint assay that is validated for high-throughput screening (HTS) of chemical libraries.

引用

页码：303 / 320

页数：18

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