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拟南芥细胞分裂素糖基转移酶UGT76C2的N端亮氨酸替换对酶活性的影响 被引量：2

Effect of the N-Terminal Leucine Substitution on the Enzyme Activity of Cytokinin Glucosyltransferase UGT76C2 of Arabidopsis thaliana
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摘要 UGT76C2是负责细胞分裂素N-糖基化修饰的糖基转移酶,该基因对于维持植物体内细胞分裂素动态平衡有重要作用。为了进一步研究UGT76C2酶蛋白结构与催化活性的关系,本文采用定点突变方法,将UGT76C2的N端第31位的保守亮氨酸替换为组氨酸。结果发现,突变型UGT76C2在离体实验中完全丧失了对细胞分裂素的糖基化修饰活性,该突变基因的过表达转基因植物出现与UGT76C2突变体类似的表型,转基因植物体内的两类主要细胞分裂素的N-糖苷含量显著降低。实验结果证明了UGT76C2 N端亮氨酸残基对于糖基化修饰活性的重要性。 UGT76C2 is a glucosyltransferases responsible for the N-glucosylation of cytokinins, which is important for in vivo cytokinin homeostasis. In order to study the effects of protein structure on catalytic activity, the conserved N-terminal amino acid, leucine（L31）, of UGT76C2 was replaced by histidine via site-specifi c mutation method. It was found that the mutated UGT76C2 completely lost in vitro glucosylating activity for cytokinins. The transgenic Arabidopsis plants overexpressing mutated UGT76C2 displayed a typical phonotype for ugt76c2 mutants. Moreover, the contents of N-glucosides of main cytokinins in Arabidopsis were substantially decreased. Our results demonstrated that the conserved N-terminal leucine of UGT76C2 was essential for cytokinin glucosylating activity.
作者 于惠敏 王会勇 王军 侯丙凯
机构地区 齐鲁师范学院生命科学学院 山东大学生命科学学院
出处 《植物生理学报》 CAS CSCD 北大核心 2014年第11期1659-1668,共10页 Plant Physiology Journal
基金 国家自然科学基金项目(30770214) 教育部博士学科点专项基金项目(20120131110023) 植物细胞工程与种质创新教育部重点实验室开放课题(2012-01)
关键词 糖基转移酶 定点突变 酶活性 细胞分裂素 拟南芥 glucosyltransferase site-specifi c mutation enzyme activity cytokinin Arabidopsis thaliana
分类号 Q946 [生物学—植物学]
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同被引文献34

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植物生理学报
2014年 第11期

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