Alan W L Law, Julien Lescar, Quan Hao, Masayo Kotaka
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引用次数: 0
摘要
腺苷酸激酶(AK)是一种磷酸转移酶,可催化腺嘌呤核苷酸的相互转化,从而在能量代谢中发挥重要作用。在恶性疟原虫中,已经发现了三种 AK 同工酶,即 PfAK1、PfAK2 和 GTP:AMP 磷酸转移酶(PfGAK)。PfAK1 和 PfAK2 催化 ATP 和 AMP 转化为两分子 ADP,而 PfGAK 则表现出对 GTP 和 AMP 的底物偏好,不接受 ATP 作为底物。PfGAK 在大肠杆菌中克隆和表达,并用两步层析法纯化。获得了 PfGAK 的棕色六方晶体,并进行了初步的衍射分析。对单个 PfGAK 晶体的 X 射线衍射数据进行了处理,分辨率为 2.9 Å,空间群为 P3(1)21 或 P3(2)21,单位晶胞参数为 a = b = 123.49,c = 180.82 Å,α = β = 90,γ = 120°。
Expression, purification, crystallization and preliminary X-ray analysis of Plasmodium falciparum GTP:AMP phosphotransferase.
Adenylate kinases (AKs) are phosphotransferase enzymes that catalyze the interconversion of adenine nucleotides, thereby playing an important role in energy metabolism. In Plasmodium falciparum, three AK isoforms, namely PfAK1, PfAK2 and GTP:AMP phosphotransferase (PfGAK), have been identified. While PfAK1 and PfAK2 catalyse the conversion of ATP and AMP to two molecules of ADP, PfGAK exhibits a substrate preference for GTP and AMP and does not accept ATP as a substrate. PfGAK was cloned and expressed in Escherichia coli and purified using two-step chromatography. Brown hexagonal crystals of PfGAK were obtained and a preliminary diffraction analysis was performed. X-ray diffraction data for a single PfGAK crystal were processed to 2.9 Å resolution in space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 123.49, c = 180.82 Å, α = β = 90, γ = 120°.
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