Identification of damaged proteins in human serum using modified Ehrlich's reagent to target protein-bound pyrroles

Fiona Margaret Campbell, Garry Jonathan Rucklidge, Martin David Reid, Louise Laura Cantlay, Simon P. Robins

Research output: Contribution to journalArticle

2 Citations (Scopus)
3 Downloads (Pure)

Abstract

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.

Original languageEnglish
Pages (from-to)76-82
Number of pages7
JournalAnalytical Biochemistry
Volume398
Issue number1
Early online date20 Nov 2009
DOIs
Publication statusPublished - 1 Mar 2010

Keywords

  • protein oxidation
  • protein-bound pyrroles
  • modified Ehrlich's reagent
  • detection of damaged proteins
  • proteomics
  • tail tendon collagen
  • advanced glycation
  • Maillard Reaction
  • cross-linking
  • oxidative stress
  • diabetic-rats
  • human plasma
  • lysine
  • 4-ketoaldehydes
  • model

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