The immunological and chemical detection of N-(hexanoyl)phosphatidylethanolamine and N-(hexanoyl)phosphatidylserine in an oxidative model induced by carbon tetrachloride.
Lipid peroxidation products have a high reactivity against the primary amino groups of biomolecules such as aminophospholipids, proteins, and DNA. Until now, many papers have reported about the modification of biomolecules derived from lipid peroxides. Our group has also reported that aminophospholipids, such as phosphatidylethanolamine (PE), can be modified by lipid peroxidation including 13-hydroperoxyoctadecadienoic acid (13-HPODE). The aim of this study was to examine the oxidative stress in vivo by detecting the formation of N-(hexanoyl)phosphatidylethanolamine (HEPE) and N-(hexanoyl)phosphatidylserine (HEPS), a novel hexanoyl adduct, using a liquid chromatography/tandem mass spectrometry (LC/MS/MS) and a monoclonal antibody. Consequently, we observed that the formation of HEPE and HEPS occurred in the red blood cell (RBC) ghosts modified by 13-HPODE and the oxidative stress model induced by carbon tetrachloride (CCl(4)) using LC/MS/MS monitoring hexanoyl ethanolamine (HEEA), a head group of HEPE, and hexanoyl serine (HESE) as a part of HEPS. Furthermore, we obtained a novel type of monoclonal antibody against HEPE. This antibody could recognize HEPE in the liver of rats with oxidative stress in vivo.
Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
Shinsuke
S
Hisaka
Naomi
N
Yamada
Kentaro
K
Naito
Toshihiko
T
Osawa
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002251:Carbon Tetrachloride / Q000633:toxicity; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015227:Lipid Peroxidation; D008297:Male; D023421:Models, Animal; D018384:Oxidative Stress; D010714:Phosphatidylethanolamines / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D010718:Phosphatidylserines / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D051381:Rats; D017208:Rats, Wistar; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis / Q000737:chemistry
D000911:Antibodies, Monoclonal; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D017392:Thiobarbituric Acid Reactive Substances; D002251:Carbon Tetrachloride