拟南芥血红蛋白1(AtGLB1)与过氧化氢的相互作用

Interaction of AtGLB1 and H_2O_2

拟南芥的血红蛋白1(AtGLB1)属于非共生的血红蛋白。在低氧胁迫中对植物细胞中过氧化氢(H2O2)内稳态的维持起了很重要的作用。为了检测AtGLB1与H2O2能否直接相互作用,我们扩增了拟南芥的AtGLB1基因,并将其克隆到原核表达质粒pET32a中,测序鉴定正确后转化大肠杆菌BL21。IPTG诱导目的蛋白表达后,镍离子亲和层析柱(Ni2+-NTA)纯化了靶蛋白。体外表达的氧合的AtGLB1能与H2O2直接相互作用。因此,与H2O2反应可能是AtGLB1清除低氧胁迫下产生的H2O2的一种方式。

Arabidopsis hemoglobin 1 （AtGLB1） is a kind of non -symbiotic hemoglobin. It plays an important role in sustaining H2O2 homeostasis when plant is exposed to hypoxic stress. To detect interaction of AtGLB1 and H2O2, the gene encoding AtGLB1 was amplified by PCR, and inserted into prokaryotic expression plasmid pET32a. Then the recombinant expression plasmid pET32a/AtGLB1 was identified by sequencing and transferred into E. coli BL21. The recombinant protein was expressed under the induction of IPTG, and purified by Ni^2＋ - NTA affinity chromatography column. The recombinant protein could react with H2O2 directly. Therefore, reaction with H2O2 might be one of ways that AtGLB1 scavenges H2O2 produced during hypoxia stress.