Wilson K, Mole D J, Binnie M, Homer N Z M, Zheng X, Yard B A, Iredale J P, Auer M, Webster S P
Drug Discovery Core, University/BHF Centre for Cardiovascular Science, Queen's Medical Research Institute, The University of Edinburgh, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
MRC Centre for Inflammation Research, Queen's Medical Research Institute, The University of Edinburgh, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Protein Expr Purif. 2014 Mar;95(100):96-103. doi: 10.1016/j.pep.2013.11.015. Epub 2013 Dec 4.
Kynurenine 3-monooxygenase (KMO) is an enzyme central to the kynurenine pathway of tryptophan metabolism. KMO has been implicated as a therapeutic target in several disease states, including Huntington's disease. Recombinant human KMO protein production is challenging due to the presence of transmembrane domains, which localise KMO to the outer mitochondrial membrane and render KMO insoluble in many in vitro expression systems. Efficient bacterial expression of human KMO would accelerate drug development of KMO inhibitors but until now this has not been achieved. Here we report the first successful bacterial (Escherichia coli) expression of active FLAG™-tagged human KMO enzyme expressed in the soluble fraction and progress towards its purification.
犬尿氨酸3-单加氧酶（KMO）是色氨酸代谢的犬尿氨酸途径中的一种关键酶。KMO已被认为是包括亨廷顿舞蹈症在内的多种疾病状态下的治疗靶点。由于存在跨膜结构域，重组人KMO蛋白的生产具有挑战性，这些跨膜结构域将KMO定位于线粒体外膜，并使KMO在许多体外表达系统中不溶。人KMO在细菌中的高效表达将加速KMO抑制剂的药物开发，但迄今为止尚未实现。在此，我们报告了首个在细菌（大肠杆菌）中成功表达可溶性部分的带有FLAG™标签的活性人KMO酶，并在其纯化方面取得了进展。
