Induction of Redox Instability of Bovine Myoglobin by Adduction with 4-Hydroxy-2-nonenal
- 26 March 2003
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 42 (15), 4398-4405
- https://doi.org/10.1021/bi0271695
Abstract
The redox stability of myoglobin (Mb) is compromised by many factors, including lipid oxidation and its products. 4-Hydroxy-2-nonenal (HNE) is an α,β-unsaturated aldehyde derived from the oxidation of ω-6 polyunsaturated fatty acids and is highly reactive and cytotoxic. Our objective was to study potential binding of HNE to Mb and determine how it affects redox stability. OxyMb (0.15 mM) was incubated with HNE (1 mM) at 4, 25, and 37 °C at pH 7.4 or 5.6. Samples were analyzed for MetMb formation and by Western blot analyses, LC−MS, LC−MS−MS, circular dichroism (CD), and differential scanning calorimetry (DSC). MetMb formation increased with increasing temperature and was greater at pH 5.6 than at pH 7.4 (P < 0.05). At 37 °C, HNE accelerated oxidation at pH 7.4 but not at pH 5.6 (P < 0.05). At both 25 and 4 °C, HNE accelerated oxidation at pH 7.4 and 5.6 (P < 0.05). LC−MS revealed the covalent binding of HNE to Mb at both pH values via Michael addition, while Western blot analysis indicated that HNE was bound to histidine (HIS) residues. LC−MS−MS identified six histidine residues of Mb that were readily adducted by HNE, including the proximal (HIS 93) and distal (HIS 64) histidine associated with the heme group. Secondary structure differences between control Mb and Mb incubated with HNE were not detected by CD. However, DSC revealed a decreased Tm for Mb reacted with HNE at pH 7.4, indicating Mb tertiary structure was altered in a manner consistent with destabilization. These results suggest that HNE accelerates bovine skeletal muscle OxyMb oxidation in vitro by covalent modification at histidine residues.Keywords
This publication has 15 references indexed in Scilit:
- Increased oxidative stress in Alzheimer's disease as assessed with 4-hydroxynonenal but not malondialdehydeQJM: An International Journal of Medicine, 2001
- The cytotoxic lipid peroxidation product, 4‐hydroxy‐2‐nonenal, specifically inhibits decarboxylating dehydrogenases in the matrix of plant mitochondriaFEBS Letters, 2000
- Effect of Aldehyde Lipid Oxidation Products on MyoglobinJournal of Agricultural and Food Chemistry, 2000
- Mass spectrometry and the age of the proteomeJournal of Mass Spectrometry, 1998
- First Direct Evidence for Lipid/Protein Conjugation in Oxidized Human Low Density LipoproteinPublished by Elsevier BV ,1996
- Tissue equilibration and subcellular distribution of vitamin E relative to myoglobin and lipid oxidation in displayed beefJournal of Animal Science, 1993
- Changes in Titin and Nebulin in Postmortem Bovine Muscle Revealed by Gel Electrophoresis, Western Blotting and Immunofluorescence MicroscopyJournal of Food Science, 1991
- Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydesFree Radical Biology & Medicine, 1991
- Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man.JCI Insight, 1989
- The determination of haem pigments in meatMeat Science, 1982