Analysis of oxidative modification of proteins

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Proteins are targets of oxidative modification. This unit describes detailed procedures for the analysis of popular indices of protein oxidation including protein carbonyl formation, loss of protein thiols, and nitrotyrosine and dityrosine formation, as well as isoaspartate formation. Procedures are detailed for the analysis of protein carbonyls labeled with 2,4-dinitrophenylhydrazine, tritiated sodium borohydride, and biotin-hydrazide, followed by detection measurements that are based on the distinguishing feature of each labeling chemical. Methods are outlined for the determination of protein cysteine oxidation by quantifying the loss of free protein thiols using radiolabeled [14C]-iodoacetamide. Protocols are described for the measurement of protein dityrosine by gas chromatography/ mass spectrometry, as are the details for the detection of protein nitrotyrosine by a competitive ELISA approach. Finally, methods are described for the quantification of protein-bound isoaspartate using protein-L-isoaspartyl methyltransferase that converts aberrant L-isoaspartyl residues in peptides and proteins to normal aspartyl residues.

Original languageEnglish
JournalCurrent Protocols in Protein Science
Volume2009
DOIs
StatePublished - 1 Apr 2009

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Proteins
Isoaspartic Acid
Sulfhydryl Compounds
Protein D-Aspartate-L-Isoaspartate Methyltransferase
Protein Carbonylation
Iodoacetamide
Oxidation
Gas Chromatography-Mass Spectrometry
Cysteine
Gas chromatography
Labeling
Mass spectrometry
Enzyme-Linked Immunosorbent Assay
Peptides
3-nitrotyrosine
dityrosine

Keywords

  • Carbonylation
  • Cysteine
  • Dityrosine
  • Isoaspartate
  • Nitrotyrosine
  • Protein oxidative modification

Cite this

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Analysis of oxidative modification of proteins. / Yan, Liang Jun.

In: Current Protocols in Protein Science, Vol. 2009, 01.04.2009.

Research output: Contribution to journalArticle

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