Laboratory of Cardiac Energetics, National Heart Lung and Blood Institute, Bethesda, Maryland 20817, USA.
Am J Physiol Heart Circ Physiol. 2012 Oct 15;303(8):H940-66. doi: 10.1152/ajpheart.00077.2012. Epub 2012 Aug 10.
It has become appreciated over the last several years that protein phosphorylation within the cardiac mitochondrial matrix and respiratory complexes is extensive. Given the importance of oxidative phosphorylation and the balance of energy metabolism in the heart, the potential regulatory effect of these classical signaling events on mitochondrial function is of interest. However, the functional impact of protein phosphorylation and the kinase/phosphatase system responsible for it are relatively unknown. Exceptions include the well-characterized pyruvate dehydrogenase and branched chain α-ketoacid dehydrogenase regulatory system. The first task of this review is to update the current status of protein phosphorylation detection primarily in the matrix and evaluate evidence linking these events with enzymatic function or protein processing. To manage the scope of this effort, we have focused on the pathways involved in energy metabolism. The high sensitivity of modern methods of detecting protein phosphorylation and the low specificity of many kinases suggests that detection of protein phosphorylation sites without information on the mole fraction of phosphorylation is difficult to interpret, especially in metabolic enzymes, and is likely irrelevant to function. However, several systems including protein translocation, adenine nucleotide translocase, cytochrome c, and complex IV protein phosphorylation have been well correlated with enzymatic function along with the classical dehydrogenase systems. The second task is to review the current understanding of the kinase/phosphatase system within the matrix. Though it is clear that protein phosphorylation occurs within the matrix, based on (32)P incorporation and quantitative mass spectrometry measures, the kinase/phosphatase system responsible for this process is ill-defined. An argument is presented that remnants of the much more labile bacterial protein phosphoryl transfer system may be present in the matrix and that the evaluation of this possibility will require the application of approaches developed for bacterial cell signaling to the mitochondria.
在过去的几年中,人们已经意识到心脏线粒体基质和呼吸复合物中的蛋白质磷酸化非常广泛。鉴于氧化磷酸化和心脏能量代谢平衡的重要性,这些经典信号事件对线粒体功能的潜在调节作用引起了人们的兴趣。然而,蛋白质磷酸化及其负责的激酶/磷酸酶系统的功能影响相对未知。例外包括丙酮酸脱氢酶和分支链α-酮酸脱氢酶调节系统。本综述的首要任务是更新目前主要在基质中检测蛋白质磷酸化的现状,并评估这些事件与酶功能或蛋白质加工之间的联系。为了管理这项工作的范围,我们专注于参与能量代谢的途径。现代检测蛋白质磷酸化的方法具有高灵敏度,而许多激酶的特异性低,这表明在没有磷酸化摩尔分数信息的情况下,检测蛋白质磷酸化位点是难以解释的,特别是在代谢酶中,并且可能与功能无关。然而,包括蛋白易位、腺嘌呤核苷酸转运酶、细胞色素 c 和复合物 IV 蛋白磷酸化在内的几个系统已经与经典脱氢酶系统一样与酶功能很好地相关。第二项任务是回顾目前对基质中激酶/磷酸酶系统的理解。尽管很明显,基于(32)P 掺入和定量质谱测量,蛋白质磷酸化发生在基质中,但负责这一过程的激酶/磷酸酶系统仍未明确定义。提出了一种观点,即更不稳定的细菌蛋白磷酸转移系统的残留物可能存在于基质中,而评估这种可能性将需要将为细菌细胞信号开发的方法应用于线粒体。