Modified sites and functional consequences of 4-oxo-2-nonenal adducts in HDL that are elevated in familial hypercholesterolemia.

Abstract

The lipid aldehyde 4-oxo-2-nonenal (ONE) is a highly reactive protein crosslinker derived from peroxidation of n-6 polyunsaturated fatty acids and generated together with 4-hydroxynonenal (HNE). Lipid peroxidation product-mediated crosslinking of proteins in high-density lipoprotein (HDL) causes HDL dysfunction and contributes to atherogenesis. Although HNE is relatively well-studied, the role of ONE in atherosclerosis and in modifying HDL is unknown. Here, we found that individuals with familial hypercholesterolemia (FH) had significantly higher ONE-ketoamide (lysine) adducts in HDL (54.6 ± 33.8 pmol/mg) than healthy controls (15.3 ± 5.6 pmol/mg). ONE crosslinked apolipoprotein A-I (apoA-I) on HDL at a concentration of > 3 mol ONE per 10 mol apoA-I (0.3 eq), which was 100-fold lower than HNE, but comparable to the potent protein crosslinker isolevuglandin. ONE-modified HDL partially inhibited HDL's ability to protect against lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNFα) and interleukin-1β (IL-1β) gene expression in murine macrophages. At 3 eq, ONE dramatically decreased apoA-I exchange from HDL, from ∼46.5 to ∼18.4% (p < 0.001). Surprisingly, ONE modification of HDL or apoA-I did not alter macrophage cholesterol efflux capacity. LC-MS/MS analysis revealed that Lys-12, Lys-23, Lys-96, and Lys-226 in apoA-I are modified by ONE ketoamide adducts. Compared with other dicarbonyl scavengers, pentylpyridoxamine (PPM) most efficaciously blocked ONE-induced protein crosslinking in HDL and also prevented HDL dysfunction in an in vitro model of inflammation. Our findings show that ONE-HDL adducts cause HDL dysfunction and are elevated in individuals with FH who have severe hypercholesterolemia.

• Authors: Linda S May-Zhang, Valery Yermalitsky, John T Melchior, Jamie Morris, Keri A Tallman, Mark S Borja, Tiffany Pleasent, Venkataraman Amarnath, Wenliang Song, Patricia G Yancey, W Sean Davidson, MacRae F Linton, Sean S Davies
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 294 Issue 50 Pg. 19022-19033 (12 13 2019) ISSN: 1083-351X [Electronic] United States
• PMID: 31666337 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2019 May-Zhang et al.

Chemical References

• 4-oxo-2-nonenal
• Aldehydes
• Apolipoprotein A-I
• Lipoproteins, HDL
• Lysine

Topics

• Aldehydes (analysis, metabolism)
• Animals
• Apolipoprotein A-I (metabolism)
• Atherosclerosis (metabolism)
• Cells, Cultured
• Female
• Humans
• Hyperlipoproteinemia Type II (blood, diagnosis, metabolism)
• Lipoproteins, HDL (metabolism)
• Lysine (metabolism)
• Male
• Mice
• Mice, Inbred C57BL
• Mice, Knockout