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PMID: 21394845 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

A mass spectrometric analysis of 4-hydroxy-2-(E)-nonenal modification of cytochrome c.

Journal of mass spectrometry : JMS ·Vol. 46 ·No. 3 ·2011-03-00 ·Pages 290-7

Tang X, Sayre LM, Tochtrop GP

Abstract

Cytochrome c is a key mitochondrial respiratory protein that is particularly susceptible to modification during oxidative stress. The nature of this susceptibility is linked to the mitochondrial membrane being rich in esterified linoleic acid, which predisposes this organelle to the formation of lipid peroxidation products such as 4-hydroxy-2-(E)-nonenal (4-HNE). To better understand the nature of cytochrome c modification by 4-HNE, we initiated an in vitro study utilizing a combination of MALDI-TOF mass spectrometry, LC-ESI-MS/MS and isotope labeling to monitor 4-HNE modification of cytochrome c under various conditions. The overwhelming reaction observed is Michael addition by Lys side-chains in addition to the modification of His 33. While the Lys-4-HNE adducts were generally observed to be reversible, the 4-HNE-His 33 was observed to be stable with half of the formed adduct surviving the denaturation and proteolysis protocols used to generate proteolytic peptides for LC-ESI-MS/MS.

MeSH Terms
Aldehydes/chemistry Amino Acid Sequence Animals Chromatography, Liquid/methods Cytochromes c/chemistry,metabolism Horses Peptide Fragments/chemistry,metabolism Protein Stability Spectrometry, Mass, Electrospray Ionization Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods Tandem Mass Spectrometry Trypsin/metabolism
Chemicals
Aldehydes Peptide Fragments Cytochromes c Trypsin 4-hydroxy-2-nonenal
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tang Xiaoxia
Department of Chemistry, Case Western Reserve University, Cleveland, OH 44106, USA.
Sayre Lawrence M
Tochtrop Gregory P
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Article Info
Journal
Journal of mass spectrometry : JMS
Abbr.
J Mass Spectrom
ISSN
1096-9888
DOI
10.1002/jms.1890
Published
2011-03-00
Pages
290-7
Language
English
Region
England
NLM ID
9504818
PMCID
PMC3903654
Subset
IM
Grants
NIA NIH HHS · R01 AG014249 · United States
NIA NIH HHS · P01 AG015885 · United States
NIA NIH HHS · AG15885 · United States
NHLBI NIH HHS · HL53315 · United States
NCI NIH HHS · R01 CA157735 · United States
NIA NIH HHS · AG14249 · United States
NHLBI NIH HHS · R01 HL053315 · United States
External Links
PubMed source DOI: 10.1002/jms.1890 PMC Full Text View journal details
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