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墙生藜中热稳定的叶绿体铜/锌超氧化物歧化酶

Heat-stable chloroplastic Cu/Zn superoxide dismutase in Chenopodium murale.

作者信息

Khanna-Chopra Renu, Sabarinath S

机构信息

Stress Physiology Laboratory, Water Technology Centre, Indian Agricultural Research Institute, New Delhi 110012, India.

出版信息

Biochem Biophys Res Commun. 2004 Aug 6;320(4):1187-92. doi: 10.1016/j.bbrc.2004.06.071.

Abstract

Chenopodium murale is a weed species having wide adaptation to different climatic regimes and experiences a temperature range of 5-45 degrees C during its life span. Higher temperatures may result in heat stress, which induces higher ROS production leading to oxidative stress in the plant. Superoxide dismutase enzyme (SOD, EC.1.15.1.1) is ubiquitous, being widely distributed among O(2)(-) consuming organisms and is the first line of defense against oxidative stress. In this study, we have characterized the thermostability of the SOD isozymes from C. murale in vitro. The leaf protein extracts, thylakoidal and stromal fractions were subjected to elevated temperatures ranging from 50 degrees C to boiling and analyzed for activity and isoform pattern of SOD. Out of six SOD isoforms, SOD V showed stability even after boiling the extract for 10min. Under high temperature treatment (>60 degrees C) there was an appearance of a new SOD band with higher electrophoretic mobility. The inhibitor studies and subcellular analysis revealed that the SOD V isoform was a chloroplastic Cu/Zn SOD. The stromal Cu/Zn SOD (SOD V) was more stable than the co-migrating thylakoidal isozyme at 80 degrees C and boiling for 10min. Hence, we report an unusual, constitutive thermostable chloroplastic Cu/Zn SOD from C. murale, which may contribute towards its heat tolerance.

摘要

墙生藜是一种适应不同气候条件的杂草物种,在其生命周期内可耐受5-45摄氏度的温度范围。较高的温度可能导致热应激,从而诱导植物产生更多的活性氧(ROS),进而引发氧化应激。超氧化物歧化酶(SOD,EC.1.15.1.1)广泛存在,在消耗O₂⁻的生物体中广泛分布,是抵御氧化应激的第一道防线。在本研究中,我们对墙生藜的SOD同工酶在体外的热稳定性进行了表征。将叶片蛋白提取物、类囊体和基质部分置于50摄氏度至沸腾的高温下处理,并分析其SOD的活性和同工型模式。在六种SOD同工型中,即使将提取物煮沸10分钟后,SOD V仍表现出稳定性。在高温处理(>60摄氏度)下,出现了一条电泳迁移率更高的新SOD条带。抑制剂研究和亚细胞分析表明,SOD V同工型是一种叶绿体铜/锌超氧化物歧化酶。在80摄氏度和煮沸10分钟的条件下,基质铜/锌超氧化物歧化酶(SOD V)比共迁移的类囊体同工酶更稳定。因此,我们报道了一种来自墙生藜的异常的、组成型热稳定叶绿体铜/锌超氧化物歧化酶,它可能有助于墙生藜的耐热性。

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