Haptoglobin binding to apolipoprotein A-I prevents damage from hydroxyl radicals on its stimulatory activity of the enzyme lecithin-cholesterol acyl-transferase.
Apolipoprotein A-I (ApoA-I), a major component of HDL, binds haptoglobin, a plasma protein transporting to liver or macrophages free Hb for preventing hydroxyl radical production. This work aimed to assess whether haptoglobin protects ApoA-I against this radical. Human ApoA-I structure, as analyzed by electrophoresis and MS, was found severely altered by hydroxyl radicals in vitro. Lower alteration of ApoA-I was found when HDL was oxidized in the presence of haptoglobin. ApoA-I oxidation was limited also when the complex of haptoglobin with both high-density lipoprotein and Hb, immobilized on resin beads, was exposed to hydroxyl radicals. ApoA-I function to stimulate cholesterol esterification was assayed in vitro by using ApoA-I-containing liposomes. Decreased stimulation was observed when liposomes oxidized without haptoglobin were used. Conversely, after oxidative stress in the presence of haptoglobin (0.5 microM monomer), the liposome activity did not change. Plasma of carrageenan-treated mice was analyzed by ELISA for the levels of haptoglobin and ApoA-I, and used to isolate HDL for MS analysis. Hydroxyproline-containing fragments of ApoA-I were found associated with low levels of haptoglobin (18 microM monomer), whereas they were not detected when the haptoglobin level increased (34-70 microM monomer). Therefore haptoglobin, when circulating at enhanced levels with free Hb during the acute phase of inflammation, might protect ApoA-I structure and function against hydroxyl radicals.
Salvatore
Alfonso
A
Dipartimento delle Scienze Biologiche, Università di Napoli Federico II, via Mezzocannone 8, 80134 Napoli, Italy.
Cigliano
Luisa
L
Bucci
Enrico M
EM
Corpillo
Davide
D
Velasco
Silvia
S
Carlucci
Alessandro
A
Pedone
Carlo
C
Abrescia
Paolo
P
D000818:Animals; D016632:Apolipoprotein A-I / Q000097:blood / Q000378:metabolism*; D053304:Apolipoprotein C-II / Q000097:blood / Q000378:metabolism; D053399:Apolipoproteins D / Q000097:blood / Q000378:metabolism; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006242:Haptoglobins / Q000378:metabolism*; D006801:Humans; D017665:Hydroxyl Radical / Q000378:metabolism*; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000097:blood / Q000378:metabolism; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism*; D011485:Protein Binding; D053719:Tandem Mass Spectrometry; D013997:Time Factors