Rat and human HARE/stabilin-2 are clearance receptors for high- and low-molecular-weight heparins

Am J Physiol Gastrointest Liver Physiol. 2009 Jun;296(6):G1191-9. doi: 10.1152/ajpgi.90717.2008. Epub 2009 Apr 9.

Abstract

The human hyaluronic acid (HA) receptor for endocytosis (HARE/stabilin-2) is the primary clearance receptor for systemic HA, chondroitin sulfates, and heparin, but not for heparan sulfate or keratan sulfate (Harris EN, Weigel JA, Weigel PH. J Biol Chem 283: 17341-17350, 2008). HARE is expressed in the sinusoidal endothelial cells (SECs) of liver and lymph nodes where it acts as a scavenger for uptake and degradation of glycosaminoglycans, both as free chains and proteoglycan fragments. Unfractionated heparin (UFH; approximately 14 kDa) and low-molecular-weight heparin (LMWH; approximately 4 kDa) are commonly used in treatments for thrombosis and cancer and in surgical and dialysis procedures. The reported half-lives of UFH and LMWH in the blood are approximately 1 h and 2-6 h, respectively. In this study, we demonstrate that anti-HARE antibodies specifically block the uptake of LMWH and UFH by isolated rat liver SECs and by human 293 cells expressing recombinant human HARE (hHARE). hHARE has a significant affinity (K(d) = 10 microM) for LMWH, and higher affinity (K(d) = 0.06 microM) for the larger UFH. Rat liver SECs or cells expressing the recombinant 190-kDa HARE isoform internalized both UFH and LMWH, and both heparins cross-compete with each other, suggesting that they share the same binding sites. These cellular results were confirmed in ELISA-like assays using purified soluble 190-hHARE ectodomain. We conclude that both UFH and LMWH are cleared by HARE/Stab2 and that the differences in the affinities of HARE binding to LMWH and UFH likely explain the longer in vivo circulating half-life of LMWH compared with UFH.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacology
  • Binding, Competitive
  • Cell Adhesion Molecules, Neuronal / physiology*
  • Cell Line
  • Dextran Sulfate / metabolism
  • Endocytosis / drug effects
  • Endocytosis / physiology
  • Endothelial Cells / metabolism
  • Enoxaparin / chemistry
  • Enoxaparin / metabolism
  • Heparin / chemistry
  • Heparin / metabolism*
  • Heparin, Low-Molecular-Weight / chemistry
  • Heparin, Low-Molecular-Weight / metabolism*
  • Humans
  • Hyaluronan Receptors / physiology*
  • Light
  • Lipoproteins, LDL / metabolism
  • Liver / cytology
  • Molecular Weight
  • Protein Binding
  • Protein Isoforms / immunology
  • Protein Isoforms / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Cell Surface / physiology*
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Scattering, Radiation
  • Transfection

Substances

  • Antibodies, Monoclonal
  • Cell Adhesion Molecules, Neuronal
  • Enoxaparin
  • Heparin, Low-Molecular-Weight
  • Hyaluronan Receptors
  • Lipoproteins, LDL
  • Protein Isoforms
  • Receptors, Cell Surface
  • Recombinant Proteins
  • STAB2 protein, human
  • acetyl-LDL
  • heparin receptor
  • hyaluronan receptor 175-kDa, rat
  • Heparin
  • Dextran Sulfate