Department of Pharmacology and Neuroscience, and the Institute for Aging and Alzheimer's Disease Research, University of North Texas Health Science Center, Fort Worth, TX 76107, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 May 15;879(17-18):1308-15. doi: 10.1016/j.jchromb.2010.08.004. Epub 2010 Aug 7.
Protein carbonylation is a major form of protein oxidation and is widely used as an indicator of oxidative stress. Carbonyl groups do not have distinguishing UV or visible, spectrophotometric absorbance/fluorescence characteristics and thus their detection and quantification can only be achieved using specific chemical probes. In this paper, we review the advantages and disadvantages of several chemical probes that have been and are still being used for protein carbonyl analysis. These probes include 2,4-dinitrophenylhydazine (DNPH), tritiated sodium borohydride ([(3)H]NaBH(4)), biotin-containing probes, and fluorescence probes. As our discussions lean toward gel-based approaches, utilizations of these probes in 2D gel-based proteomic analysis of carbonylated proteins are illustrated where applicable. Analysis of carbonylated proteins by ELISA, immunofluorescent imaging, near infrared fluorescence detection, and gel-free proteomic approaches are also discussed where appropriate. Additionally, potential applications of blue native gel electrophoresis as a tool for first dimensional separation in 2D gel-based analysis of carbonylated proteins are discussed as well.
蛋白质羰基化是蛋白质氧化的主要形式，被广泛用作氧化应激的指标。羰基基团没有独特的紫外或可见分光光度吸收/荧光特性，因此只能使用特定的化学探针来检测和定量。本文综述了几种已被和仍在用于蛋白质羰基分析的化学探针的优缺点。这些探针包括 2,4-二硝基苯肼（DNPH）、氚化钠硼氢化物([(3)H]NaBH4)、含生物素的探针和荧光探针。由于我们的讨论倾向于基于凝胶的方法，因此在适用的情况下，举例说明了这些探针在基于二维凝胶的羰基化蛋白质蛋白质组学分析中的应用。在适当的情况下，还讨论了通过 ELISA、免疫荧光成像、近红外荧光检测和无胶蛋白质组学方法分析羰基化蛋白质的情况。此外，还讨论了蓝色天然凝胶电泳作为二维凝胶分析中羰基化蛋白质第一维分离工具的潜在应用。
