Arabidopsis 3-Deoxy-d-Arabino-Heptulosonate 7-Phosphate (DAHP) Synthases of the Shikimate Pathway Display Both Manganese- and Cobalt-Dependent Activities.

IF 2.3 3区生物学 Q2 PLANT SCIENCES

Plant Direct Pub Date : 2025-01-21 eCollection Date: 2025-01-01 DOI:10.1002/pld3.70037

Ryo Yokoyama, Hiroshi A Maeda

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引用次数: 0

Abstract

The plant shikimate pathway directs a significant portion of photosynthetically assimilated carbon into the downstream biosynthetic pathways of aromatic amino acids (AAA) and aromatic natural products. 3-Deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthase (hereafter DHS) catalyzes the first step of the shikimate pathway, playing a critical role in controlling the carbon flux from central carbon metabolism into the AAA biosynthesis. Previous biochemical studies suggested the presence of manganese- and cobalt-dependent DHS enzymes (DHS-Mn and DHS-Co, respectively) in various plant species. Unlike well-studied DHS-Mn, however, the identity of DHS-Co is still unknown. Here, we show that all three DHS isoforms of Arabidopsis thaliana exhibit both DHS-Mn and DHS-Co activities in vitro. A phylogenetic analysis of various DHS orthologs and related sequences showed that Arabidopsis 3-deoxy-D-manno-octulosonate-8-phosphate synthase (KDOPS) proteins were closely related to microbial Type I DHSs. Despite their sequence similarity, these Arabidopsis KDOPS proteins showed no DHS activity. Meanwhile, optimization of the DHS assay conditions led to the successful detection of DHS-Co activity from Arabidopsis DHS recombinant proteins. Compared with DHS-Mn, DHS-Co activity displayed the same redox dependency but distinct optimal pH and cofactor sensitivity. Our work provides biochemical evidence that the DHS isoforms of Arabidopsis possess DHS-Co activity.

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莽草酸途径的3-脱氧-d-阿拉伯-七磷酸庚糖酸酯（DAHP）合成酶显示出锰和钴依赖活性。

植物莽草酸途径将相当一部分光合同化的碳引导到下游芳香氨基酸（AAA）和芳香天然产物的生物合成途径中。3- deoxy -d-阿拉伯-七磷酸七糖酸酯（DAHP）合成酶（DHS）是莽草酸途径的第一步催化酶，在控制中心碳代谢进入AAA生物合成的碳通量中起关键作用。以前的生化研究表明，在各种植物物种中存在锰依赖性和钴依赖性DHS酶（分别为DHS- mn和DHS- co）。然而，与已得到充分研究的DHS-Mn不同，DHS-Co的身份仍然未知。在这里，我们发现拟南芥的所有三种DHS亚型在体外都具有DHS- mn和DHS- co活性。多种DHS同源物及相关序列的系统发育分析表明，拟南芥3-脱氧-d -甘露糖-八糖酸-8-磷酸合成酶（KDOPS）蛋白与微生物I型DHS密切相关。尽管序列相似，但这些拟南芥KDOPS蛋白没有DHS活性。同时，通过优化DHS检测条件，成功检测了拟南芥DHS重组蛋白的DHS- co活性。与DHS-Mn相比，DHS-Co活性表现出相同的氧化还原依赖性，但最佳pH值和辅因子敏感性不同。我们的工作提供了生化证据，证明拟南芥的DHS亚型具有DHS- co活性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Plant Direct Environmental Science-Ecology

CiteScore

5.00

自引率

3.30%

发文量

101

审稿时长

14 weeks

期刊介绍： Plant Direct is a monthly, sound science journal for the plant sciences that gives prompt and equal consideration to papers reporting work dealing with a variety of subjects. Topics include but are not limited to genetics, biochemistry, development, cell biology, biotic stress, abiotic stress, genomics, phenomics, bioinformatics, physiology, molecular biology, and evolution. A collaborative journal launched by the American Society of Plant Biologists, the Society for Experimental Biology and Wiley, Plant Direct publishes papers submitted directly to the journal as well as those referred from a select group of the societies’ journals.