The physiological role, structure, and regulation of animal alternative oxidases

所有生物都使用电子传递系统（ETS）来产生能量，以供生长和繁殖。真核生物利用线粒体中的ETS产生ATP这些ETS的成分在将电子放入链的复合物和将电子抽出的复合物方面是高度可变的。除了典型的末端氧化酶细胞色素c氧化酶（COX）外，许多生物体还含有第二种末端氧化酶，称为替代氧化酶（AOX）。虽然AOX也利用氧作为最终的电子载体并产生水，但与COX不同的是，AOX不泵送质子，也不参与产生ATP的质子动力和质子梯度。虽然乍一看这种反应似乎是能量浪费，但在许多生物体中，AOX mRNA和蛋白质水平在环境胁迫下大大增加。这意味着AOX在成功适应压力条件中发挥了作用，可能是通过赋予ETS灵活性。

All organisms use electron transport systems (ETS) to generate energy for growth and reproduction. Eukaryotic organisms use ETS in mitochondria to produce ATP and the components of these ETS are highly variable in terms of the complexes that put electrons into the chain and the complexes that pull electrons out. In addition to the typical terminal oxidase in ETS, cytochrome c oxidase (COX), many organisms contain a second terminal oxidase called alternative oxidase (AOX). While AOX also utilizes oxygen as the final electron carrier and produces water, in contrast to COX, AOX does not pump protons and does not contribute to the proton motive force and proton gradient used to generate ATP. While at first glance this reaction appears to be energetically wasteful, AOX mRNA and protein levels greatly increase during environmental stress in many organisms. This implies that AOX plays a role in the successful acclimation to stress conditions, perhaps by conferring flexibility to the ETS. AOX was initially discovered in plants and therefore most research has been conducted in this kingdom. AOX was long thought to be absent from animals, but work within the past decade has confirmed that it is present in some animals. We have a poor understanding of the taxonomic distribution of AOX in animals and know little about the in vivo expression of AOX mRNA and how it is regulated. Furthermore, little is known about AOX protein expression and post-translational regulation in animals. We propose to identify new AOX sequences in animals using a combination of bioinformatics and molecular biology. This will provide us with the data needed to better understand the evolutionary history of the enzyme. We will also be characterizing the biochemical inhibitors and activators of AOX from Crassostrea gigas (Pacific oyster) by using high-resolution respirometry in order to better understand the post-translational regulation of the animal enzyme. Finally we will be investigating the physiological role of AOX in the stress tolerance of the intertidal copepod Tigriopus californicus. We are proposing to use T. californicus as a means of studying AOX mRNA and protein expression and regulation using laboratory experiments that manipulate the environmental conditions that the animal experiences. Our work will be the first complete molecular and physiological characterization of an animal AOX and will provide a means of comparison for researchers working in other organisms.