Structural Impact of 4-Hydroxynonenal Modification on Human Cytochrome CYP4F11

Posttranslational modifications of human enzymes play a crucial role in disease development. 4-hydroxynonenal (4-HNE), a lipid peroxidation product, can modify proteins and disrupt their function. Human cytochrome CYP4F11, involved in lipid metabolism and xenobiotic degradation, was previously shown to be inhibited by 4-HNE in a malaria model, where hemozoin-induced 4-HNE formation occurs in monocytes. However, structural changes to CYP4F11 upon 4-HNE modification had not been described. In this study, we investigated these changes using differential scanning calorimetry (DSC), Fourier transform infrared (FTIR), and Raman spectroscopy. DSC thermograms revealed an increased energetic barrier to unfolding, suggesting structural reorganization. FTIR data, supported by computational analysis, showed a decrease in alpha-helix content (0.2-2.5%) and an increase in beta-structure (2.2-3.3%), along with altered disordered regions. Raman spectroscopy indicated significant changes in luminescence decay across emission wavelengths. These structural alterations induced by 4-HNE conjugation (protein lipoxidation) may significantly influence the enzymatic activity of CYP4F11, with potential implications for lipid metabolism and xenobiotic detoxification.