Servicio de Inmunología, Hospital Universitario de La Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid, Spain.
J Proteomics. 2011 Dec 21;75(2):329-38. doi: 10.1016/j.jprot.2011.09.013. Epub 2011 Sep 29.
There is increasing evidence that several reversible oxidative post-translational modifications of protein cysteines participate in cell signalling. Specific proteomic techniques are required to identify these modifications and to study their regulation in different cell processes, that are collectively known as thiol redox proteomics. Recently, fluorescence derivatization methods have been developed that enable these post-translational modifications to be studied using proteomic workflows based on two-dimensional electrophoresis, which is a relatively accessible and affordable technique. As well as enabling a large number of samples to be processed, two-dimensional electrophoresis has the advantage that it does not rely on the intensive use of mass spectrometers. This methodology allows to "visualise" redox changes in a broad context and, although identification of the modified residues is not so straightforward, complementary derivatization can overcome this drawback. Here we review the different derivatization strategies that have been employed in these studies, comparing their advantages and potential limitations. We also review the applications and results obtained, with particular emphasis on those involving (patho)physiological stimuli, thereby showing the potential of these techniques to study the thiol redox proteome.
越来越多的证据表明，蛋白质半胱氨酸的几种可还原氧化翻译后修饰参与细胞信号转导。需要特定的蛋白质组学技术来鉴定这些修饰，并研究它们在不同细胞过程中的调控，这些过程统称为硫醇氧化还原蛋白质组学。最近，已经开发出荧光衍生化方法，可使用基于二维电泳的蛋白质组学工作流程研究这些翻译后修饰，二维电泳是一种相对容易获得且经济实惠的技术。二维电泳不仅能够处理大量样本，而且还具有不依赖于质谱仪的密集使用的优点。这种方法允许在更广泛的背景下“可视化”氧化还原变化，尽管修饰残基的鉴定并不那么直接，但互补衍生化可以克服这一缺点。在这里，我们回顾了这些研究中使用的不同衍生化策略，比较了它们的优点和潜在限制。我们还回顾了应用和获得的结果，特别强调了那些涉及（病理）生理刺激的结果，从而展示了这些技术研究硫醇氧化还原蛋白质组的潜力。
