The effects of HNE on ovine oxymyoglobin redox stability in a microsome model

- The effect of hydroxynonenal (HNE) on ovine myoglobin redox stability was examined.
- HNE enhanced myoglobin oxidation at all pH/temperature conditions studied.
- Mono-, di- and tri-HNE adducts were identified in ovine myoglobin.
- Sites of adduction occurred at histidine 120, histidine 25 and histidine 65.

The effect of 4-hydroxy-2-nonenal (HNE), a secondary lipid oxidation product, on ovine myoglobin (Mb) redox stability was investigated. HNE increased oxymyoglobin (OxyMb) oxidation under all pH/temperature conditions studied. Mono-, di- and tri-HNE adducts were detected by ESI-Q-TOF MS analysis. Sites of adduction, His 120, His 25 and His 65, were determined by ESI-CID-MS/MS analysis. The relationship between ovine Mb (with/without HNE) and lipid oxidation was also studied in a microsome model in the presence of α-tocopherol. Surprisingly, preincubation of Mb with HNE did not affect subsequent Mb redox stability in the microsome model (P < 0.05). Microsomes with elevated concentrations of α-tocopherol delayed lipid and Mb oxidations relative to controls. HNE-treated ovine Mb caused greater lipid oxidation compared to control ovine Mb in control microsomes (P < 0.05). This study demonstrated an interaction between ovine Mb oxidation and lipid oxidation.