We previously showed (1) that a peptide, Ac-hE18A-NH2, in which the arginine-rich heparin-binding domain of apolipoprotein E (apoE) [residues 141-150] (LRKLRKRLLR), covalently linked to 18A (DWLKAFYDKVAEKLKEAF; a class A amphipathic helix with high lipid affinity), enhanced LDL uptake and clearance. Because VLDL and remnants contain more cholesterol per particle than LDL, enhanced hepatic clearance of VLDL could lead to an effective lowering of plasma cholesterol. Therefore, in the present article we compared the ability of this peptide to mediate/facilitate the uptake and degradation of LDL and VLDL in HepG2 cells. The peptide Ac-hE18A-NH2, but not Ac-18A-NH2, enhanced the uptake of LDL by HepG2 cells 5-fold and its degradation 2-fold, The association of the peptides with VLDL resulted in the displacement of native apoE; however, only Ac-hE18A-NH2 but not Ac-18A-NH2 caused markedly enhanced uptake (6-fold) and degradation (3-fold) of VLDL. Ac-hE18A-NH2 also enhanced the uptake (15-fold) and degradation (2-fold) of trypsinized VLDL Sf 100-400 (containing no immunodetectable apoE), indicating that the peptide restored the cellular interaction of VLDL in the absence of its essential native ligand (apoE). Pretreatment of HepG2s with heparinase and heparitinase abrogated all peptide-mediated enhanced cellular activity, implicating a role for cell-surface heparan sulfate proteoglycans (HSPG). Intravenous administration of AchE18A-NH2 into apoE gene knockout mice reduced plasma cholesterol by 88% at 6 h and 30% at 24 h after injection. We conclude that this dual-domain peptide associates with LDL and VLDL and results in rapid hepatic uptake via a HSPG-facilitated pathway.