Tyrosine 192 in apolipoprotein A-I is the major site of nitration and chlorination by myeloperoxidase, but only chlorination markedly impairs ABCA1-dependent cholesterol transport

Baohai Shao, Constanze Bergt, Xiaoyun Fu, Pattie Green, John C Voss, Michael N. Oda, John F. Oram, Jay W. Heinecke

Research output: Contribution to journalArticle

189 Citations (Scopus)

Abstract

High density lipoprotein (HDL) isolated from human atherosclerotic lesions and the blood of patients with established coronary artery disease contains elevated levels of 3-nitrotyrosine and 3-chlorotyrosine. Myeloperoxidase (MPO) is the only known source of 3-chlorotyrosine in humans, indicating that MPO oxidizes HDL in vivo. In the current studies, we used tandem mass spectrometry to identify the major sites of tyrosine oxidation when lipid-free apolipoprotein A-I (apoA-I), the major protein of HDL, was exposed to MPO or peroxynitrite (ONOO-). Tyrosine 192 was the predominant site of both nitration and chlorination by MPO and was also the major site of nitration by ONOO -. Electron paramagnetic spin resonance studies of spin-labeled apoA-I revealed that residue 192 was located in an unusually hydrophilic environment. Moreover, the environment of residue 192 became much more hydrophobic when apoA-I was incorporated into discoidal HDL, and Tyr 192 of HDL-associated apoA-I was a poor substrate for nitration by both myeloperoxidase and ONOO-, suggesting that solvent accessibility accounted in part for the reactivity of Tyr192. The ability of lipid-free apoA-I to facilitate ATP-binding cassette transporter A1 cholesterol transport was greatly reduced after chlorination by MPO. Loss of activity occurred in concert with chlorination of Tyr192. Both ONOO - and MPO nitrated Tyr192 in high yield, but unlike chlorination, nitration minimally affected the ability of apoA-I to promote cholesterol efflux from cells. Our results indicate that Tyr192 is the predominant site of nitration and chlorination when MPO or ONOO- oxidizes lipid-free apoA-I but that only chlorination markedly reduces the cholesterol efflux activity of apoA-I. This impaired biological activity of chlorinated apoA-I suggests that MPO-mediated oxidation of HDL might contribute to the link between inflammation and cardiovascular disease.

Original languageEnglish (US)
Pages (from-to)5983-5993
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number7
DOIs
StatePublished - Feb 18 2005

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Nitration
Chlorination
Halogenation
Apolipoprotein A-I
Peroxidase
Tyrosine
Cholesterol
HDL Lipoproteins
Electron Spin Resonance Spectroscopy
Lipids
Oxidation
Peroxynitrous Acid
ATP-Binding Cassette Transporters
Tandem Mass Spectrometry
Bioactivity
Mass spectrometry
Coronary Artery Disease
Blood
Cardiovascular Diseases
Inflammation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tyrosine 192 in apolipoprotein A-I is the major site of nitration and chlorination by myeloperoxidase, but only chlorination markedly impairs ABCA1-dependent cholesterol transport. / Shao, Baohai; Bergt, Constanze; Fu, Xiaoyun; Green, Pattie; Voss, John C; Oda, Michael N.; Oram, John F.; Heinecke, Jay W.

In: Journal of Biological Chemistry, Vol. 280, No. 7, 18.02.2005, p. 5983-5993.

Research output: Contribution to journalArticle

Shao, Baohai ; Bergt, Constanze ; Fu, Xiaoyun ; Green, Pattie ; Voss, John C ; Oda, Michael N. ; Oram, John F. ; Heinecke, Jay W. / Tyrosine 192 in apolipoprotein A-I is the major site of nitration and chlorination by myeloperoxidase, but only chlorination markedly impairs ABCA1-dependent cholesterol transport. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 7. pp. 5983-5993.
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abstract = "High density lipoprotein (HDL) isolated from human atherosclerotic lesions and the blood of patients with established coronary artery disease contains elevated levels of 3-nitrotyrosine and 3-chlorotyrosine. Myeloperoxidase (MPO) is the only known source of 3-chlorotyrosine in humans, indicating that MPO oxidizes HDL in vivo. In the current studies, we used tandem mass spectrometry to identify the major sites of tyrosine oxidation when lipid-free apolipoprotein A-I (apoA-I), the major protein of HDL, was exposed to MPO or peroxynitrite (ONOO-). Tyrosine 192 was the predominant site of both nitration and chlorination by MPO and was also the major site of nitration by ONOO -. Electron paramagnetic spin resonance studies of spin-labeled apoA-I revealed that residue 192 was located in an unusually hydrophilic environment. Moreover, the environment of residue 192 became much more hydrophobic when apoA-I was incorporated into discoidal HDL, and Tyr 192 of HDL-associated apoA-I was a poor substrate for nitration by both myeloperoxidase and ONOO-, suggesting that solvent accessibility accounted in part for the reactivity of Tyr192. The ability of lipid-free apoA-I to facilitate ATP-binding cassette transporter A1 cholesterol transport was greatly reduced after chlorination by MPO. Loss of activity occurred in concert with chlorination of Tyr192. Both ONOO - and MPO nitrated Tyr192 in high yield, but unlike chlorination, nitration minimally affected the ability of apoA-I to promote cholesterol efflux from cells. Our results indicate that Tyr192 is the predominant site of nitration and chlorination when MPO or ONOO- oxidizes lipid-free apoA-I but that only chlorination markedly reduces the cholesterol efflux activity of apoA-I. This impaired biological activity of chlorinated apoA-I suggests that MPO-mediated oxidation of HDL might contribute to the link between inflammation and cardiovascular disease.",
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T1 - Tyrosine 192 in apolipoprotein A-I is the major site of nitration and chlorination by myeloperoxidase, but only chlorination markedly impairs ABCA1-dependent cholesterol transport

AU - Shao, Baohai

AU - Bergt, Constanze

AU - Fu, Xiaoyun

AU - Green, Pattie

AU - Voss, John C

AU - Oda, Michael N.

AU - Oram, John F.

AU - Heinecke, Jay W.

PY - 2005/2/18

Y1 - 2005/2/18

N2 - High density lipoprotein (HDL) isolated from human atherosclerotic lesions and the blood of patients with established coronary artery disease contains elevated levels of 3-nitrotyrosine and 3-chlorotyrosine. Myeloperoxidase (MPO) is the only known source of 3-chlorotyrosine in humans, indicating that MPO oxidizes HDL in vivo. In the current studies, we used tandem mass spectrometry to identify the major sites of tyrosine oxidation when lipid-free apolipoprotein A-I (apoA-I), the major protein of HDL, was exposed to MPO or peroxynitrite (ONOO-). Tyrosine 192 was the predominant site of both nitration and chlorination by MPO and was also the major site of nitration by ONOO -. Electron paramagnetic spin resonance studies of spin-labeled apoA-I revealed that residue 192 was located in an unusually hydrophilic environment. Moreover, the environment of residue 192 became much more hydrophobic when apoA-I was incorporated into discoidal HDL, and Tyr 192 of HDL-associated apoA-I was a poor substrate for nitration by both myeloperoxidase and ONOO-, suggesting that solvent accessibility accounted in part for the reactivity of Tyr192. The ability of lipid-free apoA-I to facilitate ATP-binding cassette transporter A1 cholesterol transport was greatly reduced after chlorination by MPO. Loss of activity occurred in concert with chlorination of Tyr192. Both ONOO - and MPO nitrated Tyr192 in high yield, but unlike chlorination, nitration minimally affected the ability of apoA-I to promote cholesterol efflux from cells. Our results indicate that Tyr192 is the predominant site of nitration and chlorination when MPO or ONOO- oxidizes lipid-free apoA-I but that only chlorination markedly reduces the cholesterol efflux activity of apoA-I. This impaired biological activity of chlorinated apoA-I suggests that MPO-mediated oxidation of HDL might contribute to the link between inflammation and cardiovascular disease.

AB - High density lipoprotein (HDL) isolated from human atherosclerotic lesions and the blood of patients with established coronary artery disease contains elevated levels of 3-nitrotyrosine and 3-chlorotyrosine. Myeloperoxidase (MPO) is the only known source of 3-chlorotyrosine in humans, indicating that MPO oxidizes HDL in vivo. In the current studies, we used tandem mass spectrometry to identify the major sites of tyrosine oxidation when lipid-free apolipoprotein A-I (apoA-I), the major protein of HDL, was exposed to MPO or peroxynitrite (ONOO-). Tyrosine 192 was the predominant site of both nitration and chlorination by MPO and was also the major site of nitration by ONOO -. Electron paramagnetic spin resonance studies of spin-labeled apoA-I revealed that residue 192 was located in an unusually hydrophilic environment. Moreover, the environment of residue 192 became much more hydrophobic when apoA-I was incorporated into discoidal HDL, and Tyr 192 of HDL-associated apoA-I was a poor substrate for nitration by both myeloperoxidase and ONOO-, suggesting that solvent accessibility accounted in part for the reactivity of Tyr192. The ability of lipid-free apoA-I to facilitate ATP-binding cassette transporter A1 cholesterol transport was greatly reduced after chlorination by MPO. Loss of activity occurred in concert with chlorination of Tyr192. Both ONOO - and MPO nitrated Tyr192 in high yield, but unlike chlorination, nitration minimally affected the ability of apoA-I to promote cholesterol efflux from cells. Our results indicate that Tyr192 is the predominant site of nitration and chlorination when MPO or ONOO- oxidizes lipid-free apoA-I but that only chlorination markedly reduces the cholesterol efflux activity of apoA-I. This impaired biological activity of chlorinated apoA-I suggests that MPO-mediated oxidation of HDL might contribute to the link between inflammation and cardiovascular disease.

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