title: A method for detection of 4-hydroxy-2-nonenal adducts in proteins.
abstract: We developed a procedure to measure 4-hydroxy-2-nonenal (HNE)-amino acid adducts using the fluorescent probe 2-aminopyridine (2-AP). The method is based on the fact that HNE forms Michael addition-type amino acid adducts possessing an aldehyde functionality, which upon reaction with 2-AP in the presence of NaBH₃CN can be converted to their pyridylaminated derivatives. The HNE-amino acid adducts, namely Michael addition-type HNE-cysteine, HNE-histidine, and HNE-lysine adducts, after pyridylamination were resistant to conventional acid-hydrolysis conditions for protein (6N HCl/110°C/24 h) and could be detected by HPLC with a fluorescence detector. The reductive amination-based fluorescent labeling of HNE adducts is a simple and accurate technique that may be widely used to reveal increased levels of covalently modified proteins with HNE and its related aldehydes during aging and disease.
authors: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
authors: Chika
authors: C
authors: Wakita
authors: Kazuya
authors: K
authors: Honda
authors: Takahiro
authors: T
authors: Shibata
authors: Mitsugu
authors: M
authors: Akagawa
authors: Koji
authors: K
authors: Uchida
mesh_terms: D000447:Aldehydes / Q000032:analysis* / Q000737:chemistry; D000631:Aminopyridines / Q000737:chemistry; D001894:Borohydrides / Q000737:chemistry; D002623:Chemistry Techniques, Analytical; D002851:Chromatography, High Pressure Liquid; D011506:Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D000447:Aldehydes; D000631:Aminopyridines; D001894:Borohydrides; D011506:Proteins; C009282:sodium cyanoborohydride; C027576:4-hydroxy-2-nonenal; C032439:alpha-aminopyridine