The impact of glycation on apolipoprotein A-I structure and its ability to activate lecithin: cholesterol acyltransferase

Research output: Contribution to journalJournal articleResearchpeer-review

  • E. Nobecourt
  • Davies, Michael J.
  • B. E. Brown
  • L. K. Curtiss
  • D. J. Bonnet
  • F. Charlton
  • A. S. Januszewski
  • A. J. Jenkins
  • P. J. Barter
  • K. -A. Rye

Aims/hypothesis Hyperglycaemia, one of the main features of diabetes, results in non-enzymatic glycation of plasma proteins, including apolipoprotein A-I (apoA-I), the most abundant apolipoprotein in HDL. The aim of this study was to determine how glycation affects the structure of apoA-I and its ability to activate lecithin:cholesterol acyltransferase (LCAT), a key enzyme in reverse cholesterol transport.

Materials and methods Discoidal reconstituted HDL (rHDL) containing phosphatidylcholine and apoA-I ([A-I]rHDL) were prepared by the cholate dialysis method and glycated by incubation with methylglyoxal. Glycation of apoA-I was quantified as the reduction in detectable arginine, lysine and tryptophan residues. Methylglyoxal-AGE adduct formation in apoA-I was assessed by immunoblotting. (A-I)rHDL size and surface charge were determined by non-denaturing gradient gel electrophoresis and agarose gel electrophoresis, respectively. The kinetics of the LCAT reaction was investigated by incubating varying concentrations of discoidal (A-I)rHDL with a constant amount of purified enzyme. The conformation of apoA-I was assessed by surface plasmon resonance.

Results Methylglyoxal-mediated modifications of the arginine, lysine and tryptophan residues in lipid-free and lipid-associated apoA-I were time- and concentration-dependent. These modifications altered the conformation of apoA-I in regions critical for LCAT activation and lipid binding. They also decreased (A-I)rHDL size and surface charge. The rate of LCAT-mediated cholesterol esterification in (A-I)rHDL varied according to the level of apoA-I glycation and progressively decreased as the extent of apoA-I glycation increased.

Conclusions/interpretation It is concluded that glycation of apoA-I may adversely affect reverse cholesterol transport in subjects with diabetes.

Original languageEnglish
Issue number3
Pages (from-to)643-653
Number of pages11
Publication statusPublished - Mar 2007

    Research areas

  • apolipoprotein A-I, diabetes, high-density lipoproteins, lecithin : cholesterol acyltransferase, non-enzymatic glycation, HIGH-DENSITY-LIPOPROTEINS, NONENZYMATIC GLYCOSYLATION, MICELLAR COMPLEXES, DIABETES-MELLITUS, METHYLGLYOXAL, PROTEINS, BINDING, VITRO, AGE, HDL

ID: 314392458