Igamberdiev Abir U, Hill Robert D
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba, Canada.
Methods Enzymol. 2008;436:379-91. doi: 10.1016/S0076-6879(08)36021-2.
Class 1 hemoglobins are ubiquitous plant proteins induced under hypoxic conditions. They bind oxygen tightly and, as oxyhemoglobin, react with nitric oxide produced under hypoxic conditions. The reactions involved in NO production and scavenging help maintain the redox and energy status of the cell. This article describes the expression of class 1 barley (Hordeum vulgare L.) hemoglobin in E. coli cells and its purification to homogeneity. Methods for investigating the properties of purified hemoglobin and for measuring its nitric oxide scavenging activity are described. A method for isolation of a plant methemoglobin reductase is also presented.
1类血红蛋白是在缺氧条件下诱导产生的普遍存在的植物蛋白。它们能紧密结合氧气，并以氧合血红蛋白的形式与缺氧条件下产生的一氧化氮发生反应。一氧化氮产生和清除过程中涉及的反应有助于维持细胞的氧化还原和能量状态。本文描述了1类大麦（Hordeum vulgare L.）血红蛋白在大肠杆菌细胞中的表达及其纯化至均一状态的过程。还介绍了研究纯化血红蛋白特性及其一氧化氮清除活性测定的方法。同时也给出了一种植物高铁血红蛋白还原酶的分离方法。
