Cheung Yuk-Yin, Allen Mark D, Bycroft Mark, Wong Kam-Bo
Department of Biochemistry, The Chinese University of Hong Kong, Hong Kong.
Acta Crystallogr D Biol Crystallogr. 2004 Jul;60(Pt 7):1308-10. doi: 10.1107/S0907444904010996. Epub 2004 Jun 22.
Acylphosphatases catalyse the hydrolysis of the carboxyl phosphate bond in metabolites such as acetyl phosphate, 1,3-bisphosphoglycerate, succinoyl phosphate and carbamoyl phosphate. In this study, acylphosphatase (91 residues) from the hyperthermophilic archaeon Pyrococcus horikoshii has been cloned, overexpressed, purified and crystallized using the sitting-drop vapour-diffusion method using sodium formate as a precipitant at 289 K. The crystals belong to space group P3(2)21, with unit-cell parameters a = b = 85.65, c = 75.51 A. The asymmetric unit contains two molecules of acylphosphatase, with a corresponding crystal volume per protein weight of 3.9 A Da(-1) and a solvent content of 68.6%. A data set diffracting to 1.6 A resolution was collected from a single crystal at 100 K.
酰基磷酸酶催化诸如乙酰磷酸、1,3 - 二磷酸甘油酸、琥珀酰磷酸和氨甲酰磷酸等代谢物中羧基磷酸键的水解。在本研究中，嗜热古菌火之神栖热袍菌（Pyrococcus horikoshii）的酰基磷酸酶（91个残基）已被克隆、过量表达、纯化，并采用坐滴气相扩散法，以甲酸钠作为沉淀剂，在289 K下进行结晶。晶体属于空间群P3(2)21，晶胞参数a = b = 85.65，c = 75.51 Å。不对称单元包含两个酰基磷酸酶分子，每蛋白质重量对应的晶体体积为3.9 Å Da⁻¹，溶剂含量为68.6%。在100 K下从单晶收集到了分辨率为1.6 Å的数据集。
