Serum amyloid A binding to CLA-1 (CD36 and LIMPII analogous-1) mediates serum amyloid A protein-induced activation of ERK1/2 and p38 mitogen-activated protein kinases

J Biol Chem. 2005 Mar 4;280(9):8031-40. doi: 10.1074/jbc.M405009200. Epub 2004 Dec 2.

Abstract

Serum amyloid A protein (SAA) is an acute-phase reactant, known to mediate pro-inflammatory cellular responses. This study reports that CLA-1 (CD36 and LIMPII Analogous-1; human orthologue of the Scavenger Receptor Class B Type I (SR-BI)) mediates SAA uptake and downstream SAA signaling. Flow cytometry experiments revealed more than a 5-fold increase of Alexa-488 SAA uptake in HeLa cells stably transfected with CLA-1. Alexa 488-HDL uptake directly correlated with SAA uptake when determined in several CLA-1 stably transfected HeLa cell clones expressing various levels of CLA-1. SAA directly binds to CLA-1 as determined by cross-linking and colocalization of anti-CLA-1 antibody with SAA. SAA was co-internalized with transferrin to the endocytic recycling compartment pointing to a potential site of SAA metabolism. Alexa-488 SAA uptake in the CLA-1-overexpressing HeLa cells, as well as in THP-1 monocyte cell line, can be efficiently blocked by unlabeled SAA, high density lipoprotein, and other CLA-1 ligands. At the same time, markedly enhanced levels of phosphorylation of the mitogen-activated protein kinases (MAPKs), ERK1/2, and p38, were observed in cells stably transfected with CLA-1 cells following SAA stimulation when compared with mock transfected cells. The levels of the SAA-induced interleukin-8 (IL-8) secretion by CLA-1-overexpressing cells also significantly exceeded (5- to 10-fold) those detected for control cells. Synthetic amphipathic peptides possessing a structural alpha-helical motif inhibited SAA-induced activation of both MAPKs and IL-8 secretion in THP-1 cells. The results of this study demonstrate for the first time that CLA-1 functions as an endocytic SAA receptor and is involved in SAA-mediated cell signaling events associated with the immune-related and inflammatory effects of SAA.

MeSH terms

  • Amino Acid Motifs
  • Binding Sites
  • Binding, Competitive
  • Blotting, Western
  • CD36 Antigens / biosynthesis*
  • Dose-Response Relationship, Drug
  • Endocytosis
  • Enzyme Inhibitors / pharmacology
  • Flow Cytometry
  • Fluorescent Dyes / pharmacology
  • HeLa Cells
  • Humans
  • Hydrazines / pharmacology
  • Interleukin-8 / metabolism
  • Ligands
  • MAP Kinase Signaling System
  • Microscopy, Confocal
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Peptides / chemistry
  • Phosphorylation
  • Protein Binding
  • Protein Structure, Secondary
  • Receptors, Immunologic / metabolism*
  • Receptors, Scavenger
  • Scavenger Receptors, Class B
  • Serum Amyloid A Protein / biosynthesis*
  • Signal Transduction
  • Time Factors
  • Transfection
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Alexa 488 hydrazide
  • CD36 Antigens
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Hydrazines
  • Interleukin-8
  • Ligands
  • Peptides
  • Receptors, Immunologic
  • Receptors, Scavenger
  • SCARB1 protein, human
  • Scavenger Receptors, Class B
  • Serum Amyloid A Protein
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • p38 Mitogen-Activated Protein Kinases