Structural and mechanistic characterization of L-histidinol phosphate phosphatase from the polymerase and histidinol phosphatase family of proteins.
Structural and mechanistic characterization of L-histidinol phosphate phosphatase from the polymerase and histidinol phosphatase family of proteins.
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DOI：
10.1021/bi301496p
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发表时间：
2013-02-12
期刊：
影响因子：
2.9
通讯作者：
Raushel FM
中科院分区：
文献类型：
--
作者：
Ghodge SV;Fedorov AA;Fedorov EV;Hillerich B;Seidel R;Almo SC;Raushel FM
L-组氨酸磷酸磷酸酶（HPP）催化L-组氨酸磷酸水解为L-组氨酸和无机磷酸，这是L-组氨酸生物合成中的倒数第二步。来自聚合酶和组氨醇磷酸酶（PHP）蛋白家族的HPP具有三核活性位点和扭曲的（β/α）7桶蛋白折叠。这组酶与酰胺水解酶超家族密切相关。提出了PHP家族HPP酶水解磷酸单酯键的机制。重组乳酸乳球菌HPP。在大肠杆菌中表达的lactis在活性位点含有铁和锌的混合物，催化效率约为103 M−1 s−1。在无铁条件下的蛋白质的表达导致在生产酶的催化效率和锌和锰的混合物中的活性位点的两个数量级的改善。溶剂同位素和粘度效应表明，质子转移步骤和产物解离步骤是不限速的。HPP的X-射线结构由硫酸盐、L-组氨酸/磷酸盐以及结合在活性位点的L-组氨酸和砷酸盐的复合物确定。这些晶体结构和变体的催化性质用于鉴定酰胺水解酶超家族内的HPP酶家族的催化和底物识别所需的结构元件。
l-Histidinol phosphate phosphatase (HPP) catalyzes the hydrolysis of L-histidinol phosphate to L-histidinol and inorganic phosphate, the penultimate step in the biosynthesis of L-histidine. HPP from the polymerase and histidinol phosphatase (PHP) family of proteins possesses a trinuclear active site and a distorted (β/α)7-barrel protein fold. This group of enzymes is closely related to the amidohydrolase superfamily of enzymes. The mechanism of phosphomonoester bond hydrolysis by the PHP family of HPP enzymes was addressed. Recombinant HPP from Lactococcus lactis subsp. lactis that was expressed in Escherichia coli contained a mixture of iron and zinc in the active site and had a catalytic efficiency of ~103 M−1 s−1. Expression of the protein under iron-free conditions resulted in the production of enzyme with a two orders of magnitude improvement in catalytic efficiency and a mixture of zinc and manganese in the active site. Solvent isotope and viscosity effects demonstrated that proton transfer steps and product dissociation steps are not rate-limiting. X-ray structures of HPP were determined with sulfate, L-histidinol/phosphate, and a complex of L-histidinol and arsenate bound in the active site. These crystal structures and the catalytic properties of variants were used to identify the structural elements required for catalysis and substrate recognition by the HPP family of enzymes within the amidohydrolase superfamily.
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影响因子：
4.2
作者：
Cheng, Nian-Sheng
通讯作者：
Cheng, Nian-Sheng
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
14.9
作者：
ASLANIDIS, C;DEJONG, PJ
通讯作者：
DEJONG, PJ
影响因子：
14.9
作者：
Aravind, L;Koonin, EV
通讯作者：
Koonin, EV
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH