Identification of damaged proteins in human serum using modified Ehrlich's reagent to target protein-bound pyrroles.
Protein-bound pyrroles are a sign of oxidative damage. Here we report a specific method for detecting pyrrole-containing proteins using biotin-labeled Ehrlich's reagent (ER-B). After treatment of either human serum or isolated human serum proteins with various oxidizing agents, damaged, biotin-labeled components could be detected by blotting. Combining the use of ER-B with proteomic techniques allowed human serum proteins susceptible to oxidative damage to be detected and then identified by LC/MS/MS. Identification of such proteins in different human conditions such as obesity, diabetes, and cardiovascular disease should lead to the discovery of new biomarkers and the development of specific assays to monitor health status.
Campbell
Fiona M
FM
University of Aberdeen, Rowett Institute of Nutrition and Health, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, Scotland, UK. fiona.campbell@abdn.ac.uk
Rucklidge
Garry J
GJ
Reid
Martin D
MD
Cantlay
Louise
L
Robins
Simon P
SP
D001547:Benzaldehydes / Q000737:chemistry*; D001710:Biotin / Q000031:analogs & derivatives* / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D018384:Oxidative Stress; D040901:Proteomics / Q000379:methods*; D011758:Pyrroles / Q000032:analysis / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
D001547:Benzaldehydes; D001798:Blood Proteins; D007202:Indicators and Reagents; D011758:Pyrroles; D001710:Biotin