小热休克蛋白HSP17.4与过氧化氢酶2互作调控其酶活性

Small heat shock protein HSP17.4 interacts with catalas 2 and activates regulates its activity

【目的】为探究拟南芥小热休克蛋白HSP17.4与过氧化氢酶2的关系。【方法】构建带有组氨酸标签的小热休克蛋白HSP17.4的原核蛋白表达载体,将小热休克蛋白HSP17.4与过氧化氢酶2的重组蛋白诱导表达后进行体外免疫共沉淀试验,并利用过氧化氢酶活性试剂盒检测过氧化氢酶活力。【结果】成功构建了带有组氨酸标签的小热休克蛋白HSP17.4的蛋白表达载体,并通过体外免疫共沉淀试验发现带有组氨酸标签的小热休克蛋白HSP17.4能够将无纯化标签的过氧化氢酶2成功纯化出来,过氧化氢酶活性检测表明小热休克蛋白HSP17.4可以显著增强过氧化氢酶2活性。【结论】本研究证明小热休克蛋白HSP17.4与过氧化氢酶2存在相互作用,并且小热休克蛋白HSP17.4激活过氧化氢酶2酶活性,为进一步研究小热休克蛋白HSP17.4通过调控过氧化氢酶2参与非生物胁迫响应奠定基础。

[Objective]To explore the relationship between Arabidopsis small heat shock protein HSP17.4 and catalase CAT2. [Method]The His-HSP17.4 prokaryotic protein expression vector was constructed, and the recombinant protein of HSP17.4 and CAT2 was induced and expressed. Subsequently, the pull-down assay was carried out in vitro, and the catalase activity was tested by catalase activity kit. [Result]The expression vector of His-HSP17.4 was successfully constructed. The result of pull-down assay showed that His-HSP17.4 could successfully purify the CAT2 without purification tag. Catalase activity assay showed that the small heat shock protein HSP17.4 significantly enhanced CAT2 activity. [Conclusion]This study proves that small heat shock protein HSP17.4 interacts with CAT2 and activates CAT2, which lays a foundation for further research on the role of small heat shock protein HSP17.4 in abiotic stress response through regulating CAT2 activity.