TY - JOUR
T1 - Chemical probes for analysis of carbonylated proteins
T2 - A review
AU - Yan, Liang Jun
AU - Forster, Michael J.
N1 - Funding Information:
The work was supported by the National Institute on Aging , grant number PO1AG022550 .
PY - 2011/5/15
Y1 - 2011/5/15
N2 - 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 ([3H]NaBH4), 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.
AB - 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 ([3H]NaBH4), 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.
KW - Biotin
KW - Carbonylated
KW - Carbonylation
KW - Carbonyls
KW - Chemical probes
KW - Infrared fluorescence
KW - Oxidative stress
KW - Proteomics
KW - Tritiated sodium borohydride
UR - http://www.scopus.com/inward/record.url?scp=79955610787&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2010.08.004
DO - 10.1016/j.jchromb.2010.08.004
M3 - Review article
C2 - 20732835
AN - SCOPUS:79955610787
SN - 1570-0232
VL - 879
SP - 1308
EP - 1315
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 17-18
ER -