Apolipoprotein E-mediated binding of hypertriglyceridemic very low density lipoproteins to isolated low density lipoprotein receptors detected by ligand blotting

Spencer A. Brown, David P. Via, Antonio M. Gotto, William A. Bradley, Sandra H. Gianturco

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

HTG-VLDL1, like LDL, bind with high affinity to electrophoretically transferred, isolated LDL receptors partially purified from bovine adrenal glands. Ligand blotting techniques show that binding is calcium dependent; little or no binding of LDL or HTG-VLDL1 is observed in the presence of 10 mM EDTA. HTG-VLDL1 does not bind in the presence of 7 mM suramin, an inhibitor of LDL binding to the LDL receptor. Pretreatment of LDL with either thrombin or trypsin does not affect apoB-mediated LDL binding to the LDL receptor. ApoE-mediated binding of HTG-VLDL1 to the blotted LDL receptor is abolished or greatly decreased by thrombin treatment of HTG-VLDL1 trypsin treatment of HTG-VLDL1 abolishes binding. Reincorporation of apoE into trypsinized HTG-VLDL1 restores binding. These studies demonstrate unequivocally that HTG-VLDL1 bind to the LDL receptor, that the binding of HTG-VLDL1 to the isolated LDL receptor is mediated through the thrombin-accessible apoE, and that HTG-VLDL1 which bind via potentially dissociable apoE rather than non-tranferable apoB can be used for ligand blotting.

Original languageEnglish (US)
Pages (from-to)333-340
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume139
Issue number1
DOIs
StatePublished - Aug 29 1986

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Apolipoprotein E-mediated binding of hypertriglyceridemic very low density lipoproteins to isolated low density lipoprotein receptors detected by ligand blotting'. Together they form a unique fingerprint.

Cite this