MONOCLONAL-ANTIBODIES TO THE ALTERNATIVE OXIDASE OF HIGHER-PLANT MITOCHONDRIA
MONOCLONAL-ANTIBODIES TO THE ALTERNATIVE OXIDASE OF HIGHER-PLANT MITOCHONDRIA
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DOI：
10.1104/pp.89.4.1311
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发表时间：
1989-04-01
期刊：
影响因子：
7.4
通讯作者：
MCINTOSH, L
中科院分区：
文献类型：
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作者：
ELTHON, TE;NICKELS, RL;MCINTOSH, L
除了以细胞色素氧化酶终止的细胞色素链之外，高等植物线粒体电子传递链还包含以替代氧化酶终止的替代途径。 Sauromatum guttatum Schott 的替代氧化酶最近被鉴定为一组表观 Mr 为 37、36 和 35 千道尔顿 (kD) 的蛋白质。现在已经针对这些蛋白质制备了单克隆抗体，并命名为 AOA（结合替代氧化酶簇的所有三种蛋白质）、AOU（结合上部或 37 kD 蛋白质）和 AOL（结合下部或 36 和 35 kD 蛋白质）。所有三种抗体均与其各自的替代氧化酶蛋白结合，无论这些蛋白处于天然状态还是变性状态（如蛋白质印迹）。 AOA 和 AOU 抑制替代氧化酶活性约 49%，而 AOL 仅抑制 14% 活性。当单独与 Sepharose 4B 偶联时，所有三种单克隆树脂都能够保留整个替代氧化酶蛋白簇，表明这些蛋白以某种方式物理关联。这些单克隆能够在许多产热和非产热物种中结合类似的线粒体蛋白，这表明它们将可用于表征和纯化不同系统的替代氧化酶。单克隆-Sepharose 4B 树脂保留先前鉴定的替代氧化酶蛋白簇的能力，以及这些单克隆对替代氧化酶活性的抑制，支持了这些蛋白质在构成替代氧化酶中的作用。
The higher plant mitochondrial electron transport chain contains, in addition to the cytochrome chain which terminates with cytochrome oxidase, an alternative pathway that terminates with an alternative oxidase. The alternative oxidase of Sauromatum guttatum Schott has recently been identified as a cluster of proteins with apparent Mr of 37, 36, and 35 kilodaltons (kD). Monoclonal antibodies have now been prepared to these proteins and designated as AOA (binding all three proteins of the alternative oxidase cluster), AOU (binding the upper or 37 kD protein), and AOL (binding the lower or 36 and 35 kD proteins). All three antibodies bind to their respective alternative oxidase proteins whether the proteins are in their native or denatured states (as on protein blots). AOA and AOU inhibit alternative oxidase activity around 49%, whereas AOL inhibits activity only 14%. When coupled individually to Sepharose 4B, all three monoclonal resins were capable of retaining the entire cluster of alternative oxidase proteins, suggesting that these proteins are physically associated in some manner. The monoclonals were capable of binding similar mitochondrial proteins in a number of thermogenic and nonthermogenic species, indicating that they will be useful in characterizing and purifying the alternative oxidase of different systems. The ability of the monoclonal-Sepharose 4B resins to retain the cluster of previosly identified alternative oxidase proteins, along with the inhibition of alternative oxidase activity by these monoclonals, supports the role of these proteins in constituting the alternative oxidase.