Kumar Ganesh K, Klein Jon B
Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935, USA.
J Appl Physiol (1985). 2004 Mar;96(3):1178-86; discussion 1170-2. doi: 10.1152/japplphysiol.00818.2003.
The cellular responses to hypoxia are complex and characterized by alterations in the expression of a number of genes, including stress-related genes and corresponding proteins that are necessary to maintain homeostasis. The purpose of this article is to review previous and recent studies that have examined the changes in the expression and posttranslational modification of proteins in response to chronic sustained and intermittent forms of hypoxia. A large number of studies focused on the analysis of either the single protein or a subset of related proteins using one-dimensional gel electrophoresis to separate a complex set of proteins from solubilized tissues or cell extracts, followed by immunostaining of proteins using antibodies that are specific to either native or posttranslationally modified forms. On the other hand, only a limited number of studies have examined the global perturbations on protein expression by hypoxia using proteomics approach involving two-dimensional electrophoresis coupled with mass spectrometry. Results derived from specific protein analysis of a variety of tissues and cells showed that hypoxia, depending on the duration and severity of the stimulus, affects the level and the state of posttranslational modification of a subset of proteins that are associated with energy metabolism, stress response, cell injury, development, and apoptosis. Some of these earlier findings are further corroborated by recent studies that utilize a global proteomics approach, and, more importantly, results from these proteomics investigations on the effects of hypoxia provide new protein targets for further functional analysis. The anticipated new information stems from the analysis of expression, and posttranslational modification of these novel protein targets, along with gene expression profiles, offers exciting new opportunities to further define the mechanisms of cellular responses to hypoxia and to control more effectively the clinical consequences of prolonged or periodic lack of oxygen.
细胞对缺氧的反应是复杂的，其特征是许多基因的表达发生改变，包括维持体内平衡所必需的应激相关基因和相应蛋白质。本文的目的是回顾以往和近期的研究，这些研究探讨了蛋白质在慢性持续和间歇性缺氧状态下的表达变化和翻译后修饰。大量研究集中于使用一维凝胶电泳从溶解的组织或细胞提取物中分离复杂的蛋白质组，然后用针对天然或翻译后修饰形式的特异性抗体对蛋白质进行免疫染色，从而分析单一蛋白质或相关蛋白质亚组。另一方面，只有少数研究使用蛋白质组学方法（包括二维电泳结合质谱）研究缺氧对蛋白质表达的整体影响。对各种组织和细胞进行特定蛋白质分析的结果表明，缺氧根据刺激的持续时间和严重程度，会影响与能量代谢、应激反应、细胞损伤、发育和凋亡相关的一部分蛋白质的水平和翻译后修饰状态。最近利用整体蛋白质组学方法的研究进一步证实了一些早期发现，更重要的是，这些关于缺氧影响的蛋白质组学研究结果为进一步的功能分析提供了新的蛋白质靶点。预期的新信息来自对这些新蛋白质靶点的表达和翻译后修饰分析，连同基因表达谱，为进一步确定细胞对缺氧的反应机制以及更有效地控制长期或周期性缺氧的临床后果提供了令人兴奋的新机会。
