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Site-specific 5-hydroxytryptophan incorporation into apolipoprotein A-I impairs cholesterol efflux activity and high-density lipoprotein biogenesis.

Authors
  • Zamanian-Daryoush, Maryam1, 2
  • Gogonea, Valentin1, 2, 3
  • DiDonato, Anthony J1, 2
  • Buffa, Jennifer A1, 2
  • Choucair, Ibrahim1, 2, 3
  • Levison, Bruce S1
  • Hughes, Randall A4
  • Ellington, Andrew D5
  • Huang, Ying1, 2
  • Li, Xinmin S1, 2
  • DiDonato, Joseph A1, 2
  • Hazen, Stanley L6, 2, 7
  • 1 Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
  • 2 Center for Microbiome and Human Health, Cleveland Clinic, Cleveland, Ohio 44195.
  • 3 Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115.
  • 4 United States Army Research Laboratory South, University of Texas, Austin, Texas 78712. , (United States)
  • 5 Center for Systems and Synthetic Biology, University of Texas, Austin, Texas 78712.
  • 6 Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195 [email protected]
  • 7 Heart and Vascular Institute, Cleveland Clinic, Cleveland, Ohio 44195.
Type
Published Article
Journal
Journal of Biological Chemistry
Publisher
American Society for Biochemistry and Molecular Biology
Publication Date
Apr 10, 2020
Volume
295
Issue
15
Pages
4836–4848
Identifiers
DOI: 10.1074/jbc.RA119.012092
PMID: 32098873
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Apolipoprotein A-I (apoA-I) is the major protein constituent of high-density lipoprotein (HDL) and a target of myeloperoxidase-dependent oxidation in the artery wall. In atherosclerotic lesions, apoA-I exhibits marked oxidative modifications at multiple sites, including Trp72 Site-specific mutagenesis studies have suggested, but have not conclusively shown, that oxidative modification of Trp72 of apoA-I impairs many atheroprotective properties of this lipoprotein. Herein, we used genetic code expansion technology with an engineered Saccharomyces cerevisiae tryptophanyl tRNA-synthetase (Trp-RS):suppressor tRNA pair to insert the noncanonical amino acid 5-hydroxytryptophan (5-OHTrp) at position 72 in recombinant human apoA-I and confirmed site-specific incorporation utilizing MS. In functional characterization studies, 5-OHTrp72 apoA-I (compared with WT apoA-I) exhibited reduced ABC subfamily A member 1 (ABCA1)-dependent cholesterol acceptor activity in vitro (41.73 ± 6.57% inhibition; p < 0.01). Additionally, 5-OHTrp72 apoA-I displayed increased activation and stabilization of paraoxonase 1 (PON1) activity (μmol/min/mg) when compared with WT apoA-I and comparable PON1 activation/stabilization compared with reconstituted HDL (WT apoA-I, 1.92 ± 0.04; 5-OHTrp72 apoA-I, 2.35 ± 0.0; and HDL, 2.33 ± 0.1; p < 0.001, p < 0.001, and p < 0.001, respectively). Following injection into apoA-I-deficient mice, 5-OHTrp72 apoA-I reached plasma levels comparable with those of native apoA-I yet exhibited significantly reduced (48%; p < 0.01) lipidation and evidence of HDL biogenesis. Collectively, these findings unequivocally reveal that site-specific oxidative modification of apoA-I via 5-OHTrp at Trp72 impairs cholesterol efflux and the rate-limiting step of HDL biogenesis both in vitro and in vivo.

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