Predicting the function of 4-coumarate:CoA ligase (LJ4CL1) in Lonicera japonica.
Predicting the function of 4-coumarate:CoA ligase (LJ4CL1) in Lonicera japonica.
复制标题
预测金银花 4-Coumarate:CoA 连接酶 (LJ4CL1) 的功能
DOI：
10.3390/ijms15022386
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发表时间：
2014-02-10
影响因子：
5.6
通讯作者：
Huang L
中科院分区：
文献类型：
--
作者：
Yuan Y;Yu S;Yu J;Zhan Z;Li M;Liu G;Wang X;Huang L
4-香豆酸辅酶a连接酶（4cl）是一类参与苯丙酸衍生化合物代谢途径的必需酶；然而，仅根据保守结构域的序列相似性来识别这些重要酶的同源物和相似物仍然很困难。利用公共数据库中20种植物的序列数据和金银花（Lonicera japonica）的序列，我们确定了1252个单磷酸腺苷（AMP）依赖性合成酶/连接酶序列，并将其划分为3个系统发育分支。根据它们的分配，4cl属于四个亚群中的一个，在拟南芥和水稻中有已知的蛋白质特征。我们还定义了184个非冗余序列，编码含有GEICIRG基序的蛋白质，这些含有GEICIRG基序的蛋白质的分类分布表明它们在植物中具有独特的催化活性。我们进一步分析了它们在粳稻和粳稻中的转录水平。并选择表达最高的亚群基因进行结构和结合位点预测。结合液相色谱-质谱（LC-MS）分析，对假定的底物结合LJ4CL1保守结合位点的氨基酸残基、阿豆酸和4-香豆酸的结构研究得出结论，这一高表达蛋白群在花中可能加工4-香豆酸盐，占已知苯丙素衍生物的90%。以4-香豆酸酯为模板对纯化的LJ4CL1粗蛋白进行了活性分析，结果表明4-香豆酸酯具有较高的活性，是LJ4CL1的底物之一。
4-Coumarate:CoA ligases (4CLs) are a group of essential enzymes involved in the pathway of phenylpropanoid-derived compound metabolisms; however it is still difficult to identify orthologs and paralogs of these important enzymes just based on sequence similarity of the conserved domains. Using sequence data of 20 plant species from the public databases and sequences from Lonicera japonica, we define 1252 adenosine monophosphate (AMP)-dependent synthetase/ligase sequences and classify them into three phylogenetic clades. 4CLs are in one of the four subgroups, according to their partitioning, with known proteins characterized in A. thaliana and Oryza sativa. We also defined 184 non-redundant sequences that encode proteins containing the GEICIRG motif and the taxonomic distribution of these GEICIRG-containing proteins suggests unique catalytic activities in plants. We further analyzed their transcription levels in L. japonica and L. japonica. var. chinensis flowers and chose the highest expressed genes representing the subgroups for structure and binding site predictions. Coupled with liquid chromatography-mass spectrometry (LC-MS) analysis of the L. japonica flowers, the structural study on putative substrate binding amino acid residues, ferulate, and 4-coumaric acid of the conserved binding-site of LJ4CL1 leads to a conclusion that this highly expressed protein group in the flowers may process 4-coumarate that represents 90% of the known phenylpropanoid-derived compounds. The activity of purified crude LJ4CL1 protein was analyzed using 4-coumarate as template and high activity indicating that 4-coumarate is one of the substrates of LJ4CL1.
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影响因子：
14.9
作者：
BAIROCH, A
通讯作者：
BAIROCH, A
影响因子：
3.7
作者：
Cukovic, D;Ehlting, J;Douglas, CJ
通讯作者：
Douglas, CJ
影响因子：
6.9
作者：
AERTS, RJ;BAUMANN, TW
通讯作者：
BAUMANN, TW
影响因子：
64.8
作者：
Jaillon, Olivier;Aury, Jean-Marc;Wincker, Patrick
通讯作者：
Wincker, Patrick
影响因子：
13.8
作者：
Jeanmougin, F;Thompson, JD;Gibson, TJ
通讯作者：
Gibson, TJ