Structural and enzymatic characterization of the choline kinase LicA from Streptococcus pneumoniae.
Structural and enzymatic characterization of the choline kinase LicA from Streptococcus pneumoniae.
复制标题
肺炎链球菌胆碱激酶 LicA 的结构和酶学表征
DOI：
10.1371/journal.pone.0120467
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Chen Y
中科院分区：
文献类型：
--
作者：
Wang L;Jiang YL;Zhang JR;Zhou CZ;Chen Y
LicA在肺炎链球菌的细胞壁磷酸胆碱生物合成中起关键作用。在这里，我们确定了在1.94 nm处的脱辅基形式的LicA和两种复合形式的LicA-胆碱和LicA-AMP-MES的晶体结构，分别在2.01和1.45 nm处。整体结构采用典型的蛋白激酶样折叠，活性位点位于N-和C-末端结构域的缝隙中。这三种结构呈现出不同的活性位点，其在底物结合和产物释放时经历开-闭-开构象变化。结构分析结合诱变和酶学分析，使我们能够找出关键残基的胆碱激酶活性的LicA。此外，结构比较显示α7和α8螺旋之间的环可能调节底物特异性和催化活性。这些发现为原核胆碱激酶LicA的结构和作用机制提供了新的线索，并可能为抗肺炎球菌药物的合理设计提供指导。
LicA plays a key role in the cell-wall phosphorylcholine biosynthesis of Streptococcus pneumonia. Here we determined the crystal structures of apo-form LicA at 1.94 Å and two complex forms LicA-choline and LicA-AMP-MES, at 2.01 and 1.45 Å resolution, respectively. The overall structure adopts a canonical protein kinase-like fold, with the active site located in the crevice of the N- and C- terminal domains. The three structures present distinct poses of the active site, which undergoes an open-closed-open conformational change upon substrate binding and product release. The structure analyses combined with mutageneses and enzymatic assays enabled us to figure out the key residues for the choline kinase activity of LicA. In addition, structural comparison revealed the loop between helices α7 and α8 might modulate the substrate specificity and catalytic activity. These findings shed light on the structure and mechanism of the prokaryotic choline kinase LicA, and might direct the rational design of novel anti-pneumococcal drugs.
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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：
3.2
作者：
Damjanovic, Marlen;Kharat, Arun S.;Vollmer, Waldemar
通讯作者：
Vollmer, Waldemar
影响因子：
5.6
作者：
Malito, Enrico;Sekulic, Nikolina;Lavie, Arnon
通讯作者：
Lavie, Arnon
影响因子：
14.9
作者：
CORPET, F
通讯作者：
CORPET, F
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