College of Food and Bioengineering, Henan University of Science and Technology, Luoyang, 471023, Henan, China.
Key Laboratory of Microbial Resources Exploitation and Utilization, Henan University of Science and Technology, Luoyang, 471023, Henan, China.
Protoplasma. 2024 Jul;261(4):771-781. doi: 10.1007/s00709-024-01933-2. Epub 2024 Feb 12.
Dehydrins proteins accumulate and play important protective roles in most plants during abiotic stresses. The objective of this study was to characterize a YSK-type dehydrin gene, WDHN2, isolated from Triticum aestivum previously. In this work, wheat dehydrin WDHN2 was expressed in Escherichia coli and purified by immobilized metal affinity chromatography, which exhibited as a single band by sodium dodecyl sulfonate polyacrylamide gel electrophoresis and western blotting. We show that WDHN2 is capable of alleviating lactate dehydrogenase inactivation from heat and desiccation in vitro enzyme activity protection assay. In vivo assay of Escherichia coli viability demonstrates the enhancement of cell survival by the overexpression of WDHN2. The protein aggregation prevention assay explores that WDHN2 has a broad protective effect on the cellular proteome. The results show that WDHN2 is mainly accumulated in the nucleus and cytosol, suggesting that this dehydrin may exert its function in both cellular compartments. Our data suggest that WDHN2 acts as a chaperone molecular in vivo.
脱水蛋白在大多数植物应对非生物胁迫时积累并发挥重要的保护作用。本研究的目的是对先前从普通小麦中分离的 YSK 型脱水蛋白基因 WDHN2 进行表征。在这项工作中，小麦脱水蛋白 WDHN2 在大肠杆菌中表达，并通过固定化金属亲和层析进行纯化，经十二烷基硫酸钠聚丙烯酰胺凝胶电泳和 Western blot 检测为单一条带。我们表明，WDHN2 能够在体外酶活性保护实验中缓解热和干燥导致的乳酸脱氢酶失活。大肠杆菌活力的体内测定表明，WDHN2 的过表达增强了细胞的存活率。蛋白聚集预防实验表明，WDHN2 对细胞蛋白质组具有广泛的保护作用。结果表明，WDHN2 主要积累在细胞核和细胞质中，这表明该脱水蛋白可能在这两个细胞区室中发挥作用。我们的数据表明，WDHN2 在体内作为分子伴侣发挥作用。
