Pereira de Pádua Ricardo Augusto, Nonato Maria Cristina
Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, Avenida do Café s/n, 14040-903 Ribeirão Preto-SP, Brazil.
Acta Crystallogr F Struct Biol Commun. 2014 Jan;70(Pt 1):120-2. doi: 10.1107/S2053230X13033955. Epub 2013 Dec 24.
Human fumarase (HsFH) is a well-known citric acid cycle enzyme and is therefore a key component in energy metabolism. Genetic studies on human patients have shown that polymorphisms in the fumarase gene are responsible for diseases such as hereditary leiomyomatosis and renal cell cancer. As a first step in unravelling the molecular basis of the mechanism of fumarase deficiency in genetic disorders, the HsFH gene was cloned in pET-28a, heterologously expressed in Escherichia coli, purified by nickel-affinity chromatography and crystallized using the vapour-diffusion technique. X-ray diffraction experiments were performed at a synchrotron source and the structure was solved at 2.1 Å resolution by molecular replacement.
人延胡索酸酶（HsFH）是一种著名的柠檬酸循环酶，因此是能量代谢的关键组成部分。对人类患者的遗传学研究表明，延胡索酸酶基因的多态性与遗传性平滑肌瘤病和肾细胞癌等疾病有关。作为揭示遗传性疾病中延胡索酸酶缺乏机制的分子基础的第一步，HsFH基因被克隆到pET-28a中，在大肠杆菌中进行异源表达，通过镍亲和层析纯化，并使用气相扩散技术进行结晶。在同步辐射源上进行了X射线衍射实验，并通过分子置换以2.1 Å的分辨率解析了其结构。
