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Targets for covalent protein modification by 4‐hydroxynonenal/4‐hydroxyhexenal‐mediated carbonyl stress in the mitochondria
Author(s) -
Rauniyar Navin,
ProkaiTatrai Katalin,
Prokai Laszlo
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.858.4
Subject(s) - chemistry , mass spectrometry , tandem mass spectrometry , posttranslational modification , biochemistry , chromatography , enzyme
Protein carbonylation is associated with various neurodegenerative diseases. Characterization of selectivity of 4‐hydroxy‐2‐nonenal (HNE) and 4‐hydroxy‐2‐hexenal (HHE) modifications, major components of carbonyl stress, helps to understand its impact on protein function and/or activity, as well as its effects on downstream targets or other interacting proteins. Rat brain mitochondrial proteins modified by reactive carbonyl species representing end‐products of lipid peroxidation such as HNE and HHE were investigated using high performance liquid chromatography (LC) and tandem mass spectrometry (MS/MS). Shotgun‐based approach involving conventional data‐dependent acquisition on a hybrid linear ion trap‐Fourier transform ion cyclotron resonance mass spectrometer was employed after prior enrichment of HNE‐/HHE‐modified peptides by solid‐phase hydrazide strategy. Selective enrichment of substoichiometric levels of modified peptides from complex mixtures followed by LC‐MS/MS identified several peptides that were modified with HNE or HHE with amino acid resolution. Additionally, bioinformatics (Ingenuity Pathway Analysis) was used to identify biological pathways, networks, and functions significantly altered by such modifications. This research was supported by the grant AG025384 from the National Institutes of Health.