Immunochemical evidence supporting 2-pentylpyrrole formation on proteins exposed to 4-hydroxy-2-nonenal.
Previous model studies suggested the formation of lysine-based 2-pentylpyrroles as novel late adduction products formed upon exposure of proteins to the lipid peroxidation product 4-hydroxy-2-nonenal (HNE). Two 2-pentylpyrrole immunogens were prepared, one by treating keyhole limpet hemocyanin (KLH) directly with 4-oxononanal and the other by preformation of 6-(2-pentylpyrrol-1-yl)hexanoic acid from 6-aminocaproic acid and 4-oxononanal, followed by carbodiimide coupling to KLH. Pyrrole content and lysine modification in KLH were assayed independently. Following immunization of rabbits, antibody titer increased and plateaued over a 4 month period. The structural specificity of the IgG fractions of the antisera was evaluated through comprehensive competitive ELISA studies. These antibodies were used to verify the time-dependent appearance of the 2-pentylpyrrole epitope in protein exposed to HNE. Potential advantages of antibodies recognizing "advanced lipid peroxidation end products" over those recognizing "early" HNE adduction products are discussed.
Sayre
L M
LM
Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, USA. lms3@po.cwru.edu
Sha
W
W
Xu
G
G
Kaur
K
K
Nadkarni
D
D
Subbanagounder
G
G
Salomon
R G
RG
D000447:Aldehydes / Q000737:chemistry / Q000276:immunology / Q000633:toxicity*; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D003432:Cross-Linking Reagents; D000939:Epitopes / Q000737:chemistry; D007120:Immunochemistry; D011506:Proteins / Q000737:chemistry / Q000187:drug effects* / Q000276:immunology*; D011758:Pyrroles / Q000138:chemical synthesis* / Q000737:chemistry; D011817:Rabbits
D000447:Aldehydes; D000906:Antibodies; D003432:Cross-Linking Reagents; D000939:Epitopes; D011506:Proteins; D011758:Pyrroles; C027576:4-hydroxy-2-nonenal