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Dehydrin ERD14 activates glutathione transferase Phi9 in Arabidopsis thaliana under osmotic stress.
Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 3 ) Pub Date : 2019-12-20 , DOI: 10.1016/j.bbagen.2019.129506
Phuong N Nguyen 1 , Maria-Armineh Tossounian 2 , Denes S Kovacs 3 , Tran T Thu 3 , Benoit Stijlemans 4 , Didier Vertommen 5 , Jarne Pauwels 6 , Kris Gevaert 6 , Geert Angenon 7 , Joris Messens 2 , Peter Tompa 8
Affiliation  

BACKGROUND Fully intrinsically disordered plant dehydrin ERD14 can protect enzymes via its chaperone-like activity, but it was not formally linked with enzymes of the plant redox system yet. This is of particular interest, as the level of H2O2 in Arabidopsis plants increases during osmotic stress, which can be counteracted by overexpression of ERD14. METHODS The proteomic mass-spectrometry analysis of stressed plants was performed to find the candidates affected by ERD14. With cross-linking, microscale thermophoresis, and active-site titration kinetics, the interaction and influence of ERD14 on the function of two target proteins: glutathione transferase Phi9 and catalase was examined. RESULTS Under osmotic stress, redox enzymes, specifically the glutathione transferase Phi enzymes, are upregulated. Using microscale thermophoresis, we showed that ERD14 directly interacts with GSTF9 with a KD of ~25 μM. ERD14 activates the inactive GSTF9 molecules, protects GSTF9 from oxidation, and can also increases the activity of the enzyme. Aside from GSTF9, we found that ERD14 can also interact with catalase, an important cellular H2O2 scavenging enzyme, with a KD of ~0.13 μM, and protects it from dehydration-induced loss of activity. CONCLUSIONS We propose that fully intrinsically disordered dehydrin ERD14 might protect and even activate redox enzymes, helping plants to survive oxidative stress under dehydration conditions. GENERAL SIGNIFICANCE ERD14 has a direct effect on the activity of redox enzymes.

中文翻译:

脱水蛋白ERD14在渗透胁迫下激活拟南芥中的谷胱甘肽转移酶Phi9。

背景技术完全内在无序的植物脱水素ERD14可以通过其伴侣蛋白样活性来保护酶,但尚未与植物氧化还原系统的酶正式连接。这是特别令人感兴趣的,因为拟南芥植物中的H2O2含量在渗透胁迫期间增加,而这可以通过ERD14的过表达来抵消。方法对受胁迫的植物进行了蛋白质组质谱分析,以找到受ERD14影响的候选对象。通过交联,微尺度热泳和活性位滴定动力学,研究了ERD14对两种靶蛋白(谷胱甘肽转移酶Phi9和过氧化氢酶)功能的相互作用和影响。结果在渗透胁迫下,氧化还原酶,特别是谷胱甘肽转移酶Phi酶被上调。使用微尺度热泳,我们证明ERD14与GSTF9直接相互作用,KD约为25μM。ERD14激活无活性的GSTF9分子,保护GSTF9免受氧化,还可以增加酶的活性。除了GSTF9,我们发现ERD14还可以与过氧化氢酶(一种重要的细胞H2O2清除酶)相互作用,KD为〜0.13μM,并保护其免受脱水诱导的活性丧失的影响。结论我们认为完全内在失调的脱水素ERD14可能保护甚至激活氧化还原酶,从而帮助植物在脱水条件下抵抗氧化胁迫。一般意义ERD14对氧化还原酶的活性有直接影响。并且还可以增加酶的活性。除了GSTF9,我们发现ERD14还可以与过氧化氢酶(一种重要的细胞H2O2清除酶)相互作用,KD为〜0.13μM,并保护其免受脱水诱导的活性丧失。结论我们认为完全内在失调的脱水素ERD14可能保护甚至激活氧化还原酶,从而帮助植物在脱水条件下抵抗氧化胁迫。一般意义ERD14对氧化还原酶的活性有直接影响。并且还可以增加酶的活性。除了GSTF9,我们发现ERD14还可以与过氧化氢酶(一种重要的细胞H2O2清除酶)相互作用,KD为〜0.13μM,并保护其免受脱水诱导的活性丧失的影响。结论我们认为完全内在失调的脱水素ERD14可能保护甚至激活氧化还原酶,从而帮助植物在脱水条件下抵抗氧化胁迫。一般意义ERD14对氧化还原酶的活性有直接影响。帮助植物在脱水条件下抵抗氧化胁迫。一般意义ERD14对氧化还原酶的活性有直接影响。帮助植物在脱水条件下抵抗氧化胁迫。一般意义ERD14对氧化还原酶的活性有直接影响。
更新日期:2019-12-21
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