Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research-IPK, Corrensstrasse 3, 06466 Gatersleben, Germany.
Plant J. 2012 Sep;71(5):850-9. doi: 10.1111/j.1365-313X.2012.05035.x. Epub 2012 Jun 22.
The metabolic function of the predicted Arabidopsis tyrosine aminotransferase (TAT) encoded by the At5g53970 gene was studied using two independent knock-out mutants. Gas chromatography-mass spectrometry based metabolic profiling revealed a specific increase in tyrosine levels, supporting the proposed function of At5g53970 as a tyrosine-specific aminotransferase not involved in tyrosine biosynthesis, but rather in utilization of tyrosine for other metabolic pathways. The TAT activity of the At5g53970-encoded protein was verified by complementation of the Escherichia coli tyrosine auxotrophic mutant DL39, and in vitro activity of recombinantly expressed and purified At5g53970 was found to be specific for tyrosine. To investigate the physiological role of At5g53970, the consequences of reduction in tyrosine utilization on metabolic pathways having tyrosine as a substrate were analysed. We found that tocopherols were substantially reduced in the mutants and we conclude that At5g53970 encodes a TAT important for the synthesis of tocopherols in Arabidopsis.
使用两个独立的敲除突变体研究了由 At5g53970 基因编码的拟南芥酪氨酸氨基转移酶(TAT)的代谢功能。基于气相色谱-质谱的代谢谱分析显示酪氨酸水平特异性增加,支持 At5g53970 作为一种特异性的酪氨酸氨基转移酶的功能,不参与酪氨酸的生物合成,而是参与酪氨酸用于其他代谢途径的利用。通过对大肠杆菌酪氨酸营养缺陷型突变体 DL39 的互补,验证了由 At5g53970 编码的蛋白质的 TAT 活性,并且发现重组表达和纯化的 At5g53970 的体外活性特异性针对酪氨酸。为了研究 At5g53970 的生理作用,分析了减少酪氨酸利用对以酪氨酸为底物的代谢途径的影响。我们发现突变体中的生育酚明显减少,我们得出结论,At5g53970 编码的 TAT 对拟南芥生育酚的合成很重要。