Chemical probes for analysis of carbonylated proteins: a review.
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.
Yan
Liang-Jun
LJ
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.
Forster
Michael J
MJ
D000818:Animals; D001710:Biotin / Q000737:chemistry; D001894:Borohydrides / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D006834:Hydrazines / Q000737:chemistry; D050050:Protein Carbonylation; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*
D001894:Borohydrides; D005456:Fluorescent Dyes; D006834:Hydrazines; D011506:Proteins; C446799:dinitrophenylhydrazine; D001710:Biotin; C025364:sodium borohydride