title: Structural and functional changes in human insulin induced by the lipid peroxidation byproducts 4-hydroxy-2-nonenal and 4-hydroxy-2-hexenal.
abstract: Lipid peroxidation produces many reactive byproducts including 4-hydroxy-2-hexenal (HHE) and 4-hydroxy-2-nonenal (HNE) derived from the peroxidation of n-3 and n-6 polyunsaturated fatty acids, respectively. HNE and HHE can modify circulating biomolecules through the formation of covalent adducts. It remains, however, unknown whether HHE and HNE could induce functional and structural changes in the insulin molecule, which may in turn be pivotal in the development of insulin resistance and diabetes. Recombinant human insulin was incubated in the presence of HHE or HNE, and the formation of covalent adducts on insulin was analyzed by mass spectrometry analysis. Insulin tolerance test in mice and stimulation of glucose uptake by 3T3 adipocytes and L6 muscle cells were used to evaluate the biological efficiency of adducted insulin compared with the native one. One to 5 adducts were formed on insulin through Michael adduction, involving histidine residues. Glucose uptake in 3T3-L1 and L6C5 cells as well as the hypoglycemic effect in mice was significantly reduced after treatment with adducted insulin compared to native insulin. The formation of HNE- and HHE-Michael adducts significantly disrupts the biological activity of insulin. These structural and functional abnormalities of the insulin molecule might contribute to the pathogenesis of insulin resistance.
authors.affiliation: Université de Lyon, Oullins, France. n.pillon@gmail.com
authors.forename: Nicolas J
authors.initials: NJ
authors.lastname: Pillon
authors.forename: Roxane E
authors.initials: RE
authors.lastname: Vella
authors.forename: Laurent
authors.initials: L
authors.lastname: Souleere
authors.forename: Michel
authors.initials: M
authors.lastname: Becchi
authors.forename: Michel
authors.initials: M
authors.lastname: Lagarde
authors.forename: Christophe O
authors.initials: CO
authors.lastname: Soulage
mesh_terms: D041721:3T3-L1 Cells; D000447:Aldehydes / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006801:Humans; D007328:Insulin / Q000737:chemistry* / Q000378:metabolism*; D015227:Lipid Peroxidation; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D000447:Aldehydes; D007328:Insulin; D011994:Recombinant Proteins; C045934:4-hydroxy-2-hexenal; C027576:4-hydroxy-2-nonenal