Macrophage receptors responsible for distinct recognition of low density lipoprotein containing pyrrole or pyridinium adducts: models of oxidized low density lipoprotein

J Lipid Res. 2000 Sep;41(9):1455-63.

Abstract

Oxidation of low density lipoproteins (LDL) induced by incubation with Cu(2+) ions results in the formation of a heterogeneous group of aldehydic adducts on lysyl residues (Lys) of apolipoprotein B (apoB) that are thought to be responsible for the uptake of oxidized LDL (oxLDL) by macrophages. To define the structural and chemical criteria governing such cell recognition, we induced two modifications of lysines in LDL that mimic prototypic adducts present in oxLDL; namely, epsilon-amino charge-neutralizing pyrrolation by treatment with 2,5-hexanedione (hdLDL), and epsilon-amino charge-retaining pyridinium formation via treatment with 2,4,6-trimethylpyrylium (tmpLDL). Both modifications led to recognition by receptors on mouse peritoneal macrophages (MPM). To assess whether the murine scavenger receptor class A-I (mSR-A) was responsible for recognition of hdLDL or tmpLDL in MPM, we measured binding at 4 degrees C and degradation at 37 degrees C of these modified forms of (125)I-labeled LDL by mSR-A-transfected CHO cells. Although uptake and degradation of hdLDL by mSR-A-transfected CHO cells was quantitatively similar to that of the positive control, acLDL, tmpLDL was not recognized by these cells. However, both tmpLDL and hdLDL were recognized by 293 cells that had been transfected with CD36. In the human monocytic cell line THP-1 that had been activated with PMA, uptake of tmpLDL was significantly inhibited by blocking monoclonal antibodies to CD36, further suggesting recognition of tmpLDL by this receptor. Macrophage uptake and degradation of LDL oxidized by brief exposure to Cu(2+) was inhibited more effectively by excess tmpLDL and hdLDL than was more extensively oxidized LDL, consistent with the recognition of the former by CD36 and the latter primarily by SR-A.Collectively, these studies suggest that formation of specific pyrrole adducts on LDL leads to recognition by both the mSR-A and mouse homolog of CD36 expressed on MPM, while formation of specific pyridinium adducts on LDL leads to recognition by the mouse homolog of CD 36 but not by mSR-A. As such, these two modifications of LDL may represent useful models for dissecting the relative contributions of specific modifications on LDL produced during oxidation, to the cellular uptake of this heterogeneous ligand.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / pharmacology
  • CD36 Antigens / drug effects
  • CD36 Antigens / physiology
  • CHO Cells
  • Cell Line
  • Cricetinae
  • Female
  • Humans
  • Lipoproteins, LDL / blood
  • Lipoproteins, LDL / metabolism*
  • Macrophages, Peritoneal / physiology*
  • Mice
  • Mice, Inbred C57BL
  • Monocytes / drug effects
  • Monocytes / physiology
  • Pyridinium Compounds
  • Pyrroles
  • Receptors, Immunologic / metabolism*
  • Receptors, Scavenger
  • Scavenger Receptors, Class A
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transfection

Substances

  • Antibodies, Monoclonal
  • CD36 Antigens
  • Lipoproteins, LDL
  • MSR1 protein, human
  • Msr1 protein, mouse
  • Pyridinium Compounds
  • Pyrroles
  • Receptors, Immunologic
  • Receptors, Scavenger
  • Scavenger Receptors, Class A
  • oxidized low density lipoprotein
  • Tetradecanoylphorbol Acetate