Abstract
Apolipoprotein (apo) E mediates lipoprotein binding to cellular lipoprotein receptors. Previously we reported that a synthetic peptide representing a linear dimeric repeat of amino acids 141-155 binds cellular LDL receptors. To prepare an apoE peptide that bound to both cholesterol-rich lipoproteins and lipoprotein receptors, an NH2-terminal acetylated apoE dimer peptide was synthesized. This acetylated peptide preferentially associated with lipoproteins in plasma, whereas nonacylated peptides were poor lipid binders. Acetylated peptide/LDL complexes (molar ratios of 4-5:1) enhanced the interaction of LDL with cultured human fibroblasts by 7-12-fold. Participation by both receptors and cell surface heparin sulfate proteoglycans was observed. When a preformed peptide/125I-LDL complex was injected intravenously into C57BL/6J apoE-deficient mice, its rate of removal was threefold higher than that of 125I-LDL alone. The liver and the spleen were major tissue distribution sites. Intravenous administration of free acetylated peptide resulted in a 30% reduction in total plasma cholesterol within 3-30 min, which reflected a 40-50% and 20-26% reduction in very low density lipoproteins and intermediate density lipoproteins, respectively. Therefore, this peptide selectively associated with cholesterol-rich lipoproteins and mediated their acute clearance in vivo.
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- Bachmair A., Finley D., Varshavsky A. In vivo half-life of a protein is a function of its amino-terminal residue. Science. 1986 Oct 10;234(4773):179–186. doi: 10.1126/science.3018930. [DOI] [PubMed] [Google Scholar]
- Boisvert W. A., Spangenberg J., Curtiss L. K. Treatment of severe hypercholesterolemia in apolipoprotein E-deficient mice by bone marrow transplantation. J Clin Invest. 1995 Aug;96(2):1118–1124. doi: 10.1172/JCI118098. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brand K., Banka C. L., Mackman N., Terkeltaub R. A., Fan S. T., Curtiss L. K. Oxidized LDL enhances lipopolysaccharide-induced tissue factor expression in human adherent monocytes. Arterioscler Thromb. 1994 May;14(5):790–797. doi: 10.1161/01.atv.14.5.790. [DOI] [PubMed] [Google Scholar]
- Cardin A. D., Hirose N., Blankenship D. T., Jackson R. L., Harmony J. A., Sparrow D. A., Sparrow J. T. Binding of a high reactive heparin to human apolipoprotein E: identification of two heparin-binding domains. Biochem Biophys Res Commun. 1986 Jan 29;134(2):783–789. doi: 10.1016/s0006-291x(86)80489-2. [DOI] [PubMed] [Google Scholar]
- Curtiss L. K., Smith R. S. Localization of two epitopes of apolipoprotein A-I that are exposed on human high density lipoproteins using monoclonal antibodies and synthetic peptides. J Biol Chem. 1988 Sep 25;263(27):13779–13785. [PubMed] [Google Scholar]
- Dyer C. A., Cistola D. P., Parry G. C., Curtiss L. K. Structural features of synthetic peptides of apolipoprotein E that bind the LDL receptor. J Lipid Res. 1995 Jan;36(1):80–88. [PubMed] [Google Scholar]
- Dyer C. A., Curtiss L. K. A synthetic peptide mimic of plasma apolipoprotein E that binds the LDL receptor. J Biol Chem. 1991 Dec 5;266(34):22803–22806. [PubMed] [Google Scholar]
- Dyer C. A., Smith R. S., Curtiss L. K. Only multimers of a synthetic peptide of human apolipoprotein E are biologically active. J Biol Chem. 1991 Aug 15;266(23):15009–15015. [PubMed] [Google Scholar]
- Farndale R. W., Buttle D. J., Barrett A. J. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta. 1986 Sep 4;883(2):173–177. doi: 10.1016/0304-4165(86)90306-5. [DOI] [PubMed] [Google Scholar]
- Farndale R. W., Sayers C. A., Barrett A. J. A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res. 1982;9(4):247–248. doi: 10.3109/03008208209160269. [DOI] [PubMed] [Google Scholar]
- Gamble W., Vaughan M., Kruth H. S., Avigan J. Procedure for determination of free and total cholesterol in micro- or nanogram amounts suitable for studies with cultured cells. J Lipid Res. 1978 Nov;19(8):1068–1070. [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S. Binding and degradation of low density lipoproteins by cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. J Biol Chem. 1974 Aug 25;249(16):5153–5162. [PubMed] [Google Scholar]
- Herz J., Goldstein J. L., Strickland D. K., Ho Y. K., Brown M. S. 39-kDa protein modulates binding of ligands to low density lipoprotein receptor-related protein/alpha 2-macroglobulin receptor. J Biol Chem. 1991 Nov 5;266(31):21232–21238. [PubMed] [Google Scholar]
- Herz J., Hamann U., Rogne S., Myklebost O., Gausepohl H., Stanley K. K. Surface location and high affinity for calcium of a 500-kd liver membrane protein closely related to the LDL-receptor suggest a physiological role as lipoprotein receptor. EMBO J. 1988 Dec 20;7(13):4119–4127. doi: 10.1002/j.1460-2075.1988.tb03306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Innerarity T. L., Pitas R. E., Mahley R. W. Binding of arginine-rich (E) apoprotein after recombination with phospholipid vesicles to the low density lipoprotein receptors of fibroblasts. J Biol Chem. 1979 May 25;254(10):4186–4190. [PubMed] [Google Scholar]
- Ishibashi S., Herz J., Maeda N., Goldstein J. L., Brown M. S. The two-receptor model of lipoprotein clearance: tests of the hypothesis in "knockout" mice lacking the low density lipoprotein receptor, apolipoprotein E, or both proteins. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4431–4435. doi: 10.1073/pnas.91.10.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ji Z. S., Brecht W. J., Miranda R. D., Hussain M. M., Innerarity T. L., Mahley R. W. Role of heparan sulfate proteoglycans in the binding and uptake of apolipoprotein E-enriched remnant lipoproteins by cultured cells. J Biol Chem. 1993 May 15;268(14):10160–10167. [PubMed] [Google Scholar]
- Ji Z. S., Fazio S., Lee Y. L., Mahley R. W. Secretion-capture role for apolipoprotein E in remnant lipoprotein metabolism involving cell surface heparan sulfate proteoglycans. J Biol Chem. 1994 Jan 28;269(4):2764–2772. [PubMed] [Google Scholar]
- Kowal R. C., Herz J., Goldstein J. L., Esser V., Brown M. S. Low density lipoprotein receptor-related protein mediates uptake of cholesteryl esters derived from apoprotein E-enriched lipoproteins. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5810–5814. doi: 10.1073/pnas.86.15.5810. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langer R. New methods of drug delivery. Science. 1990 Sep 28;249(4976):1527–1533. doi: 10.1126/science.2218494. [DOI] [PubMed] [Google Scholar]
- Mahley R. W. Apolipoprotein E: cholesterol transport protein with expanding role in cell biology. Science. 1988 Apr 29;240(4852):622–630. doi: 10.1126/science.3283935. [DOI] [PubMed] [Google Scholar]
- Mahley R. W., Ji Z. S., Brecht W. J., Miranda R. D., He D. Role of heparan sulfate proteoglycans and the LDL receptor-related protein in remnant lipoprotein metabolism. Ann N Y Acad Sci. 1994 Sep 10;737:39–52. doi: 10.1111/j.1749-6632.1994.tb44300.x. [DOI] [PubMed] [Google Scholar]
- Mahley R. W., Weisgraber K. H., Hussain M. M., Greenman B., Fisher M., Vogel T., Gorecki M. Intravenous infusion of apolipoprotein E accelerates clearance of plasma lipoproteins in rabbits. J Clin Invest. 1989 Jun;83(6):2125–2130. doi: 10.1172/JCI114126. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
- Medh J. D., Fry G. L., Bowen S. L., Pladet M. W., Strickland D. K., Chappell D. A. The 39-kDa receptor-associated protein modulates lipoprotein catabolism by binding to LDL receptors. J Biol Chem. 1995 Jan 13;270(2):536–540. doi: 10.1074/jbc.270.2.536. [DOI] [PubMed] [Google Scholar]
- Mims M. P., Darnule A. T., Tovar R. W., Pownall H. J., Sparrow D. A., Sparrow J. T., Via D. P., Smith L. C. A nonexchangeable apolipoprotein E peptide that mediates binding to the low density lipoprotein receptor. J Biol Chem. 1994 Aug 12;269(32):20539–20547. [PubMed] [Google Scholar]
- Piedrahita J. A., Zhang S. H., Hagaman J. R., Oliver P. M., Maeda N. Generation of mice carrying a mutant apolipoprotein E gene inactivated by gene targeting in embryonic stem cells. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4471–4475. doi: 10.1073/pnas.89.10.4471. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pitas R. E., Innerarity T. L., Arnold K. S., Mahley R. W. Rate and equilibrium constants for binding of apo-E HDLc (a cholesterol-induced lipoprotein) and low density lipoproteins to human fibroblasts: evidence for multiple receptor binding of apo-E HDLc. Proc Natl Acad Sci U S A. 1979 May;76(5):2311–2315. doi: 10.1073/pnas.76.5.2311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pitas R. E., Innerarity T. L., Mahley R. W. Cell surface receptor binding of phospholipid . protein complexes containing different ratios of receptor-active and -inactive E apoprotein. J Biol Chem. 1980 Jun 10;255(11):5454–5460. [PubMed] [Google Scholar]
- Plump A. S., Smith J. D., Hayek T., Aalto-Setälä K., Walsh A., Verstuyft J. G., Rubin E. M., Breslow J. L. Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells. Cell. 1992 Oct 16;71(2):343–353. doi: 10.1016/0092-8674(92)90362-g. [DOI] [PubMed] [Google Scholar]
- Ponsin G., Sparrow J. T., Gotto A. M., Jr, Pownall H. J. In vivo interaction of synthetic acylated apopeptides with high density lipoproteins in rat. J Clin Invest. 1986 Feb;77(2):559–567. doi: 10.1172/JCI112337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ponsin G., Strong K., Gotto A. M., Jr, Sparrow J. T., Pownall H. J. In vitro binding of synthetic acylated lipid-associating peptides to high-density lipoproteins: effect of hydrophobicity. Biochemistry. 1984 Oct 23;23(22):5337–5342. doi: 10.1021/bi00317a036. [DOI] [PubMed] [Google Scholar]
- Richardson P. D., Davies M. J., Born G. V. Influence of plaque configuration and stress distribution on fissuring of coronary atherosclerotic plaques. Lancet. 1989 Oct 21;2(8669):941–944. doi: 10.1016/s0140-6736(89)90953-7. [DOI] [PubMed] [Google Scholar]
- Shimano H., Yamada N., Katsuki M., Yamamoto K., Gotoda T., Harada K., Shimada M., Yazaki Y. Plasma lipoprotein metabolism in transgenic mice overexpressing apolipoprotein E. Accelerated clearance of lipoproteins containing apolipoprotein B. J Clin Invest. 1992 Nov;90(5):2084–2091. doi: 10.1172/JCI116091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sparrow J. T., Sparrow D. A., Culwell A. R., Gotto A. M., Jr Apolipoprotein E: phospholipid binding studies with synthetic peptides containing the putative receptor binding region. Biochemistry. 1985 Nov 19;24(24):6984–6988. doi: 10.1021/bi00345a035. [DOI] [PubMed] [Google Scholar]
- Takahashi S., Kawarabayasi Y., Nakai T., Sakai J., Yamamoto T. Rabbit very low density lipoprotein receptor: a low density lipoprotein receptor-like protein with distinct ligand specificity. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9252–9256. doi: 10.1073/pnas.89.19.9252. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weisgraber K. H., Rall S. C., Jr, Mahley R. W., Milne R. W., Marcel Y. L., Sparrow J. T. Human apolipoprotein E. Determination of the heparin binding sites of apolipoprotein E3. J Biol Chem. 1986 Feb 15;261(5):2068–2076. [PubMed] [Google Scholar]
- Westerlund J. A., Weisgraber K. H. Discrete carboxyl-terminal segments of apolipoprotein E mediate lipoprotein association and protein oligomerization. J Biol Chem. 1993 Jul 25;268(21):15745–15750. [PubMed] [Google Scholar]
- Yamada N., Shimano H., Mokuno H., Ishibashi S., Gotohda T., Kawakami M., Watanabe Y., Akanuma Y., Murase T., Takaku F. Increased clearance of plasma cholesterol after injection of apolipoprotein E into Watanabe heritable hyperlipidemic rabbits. Proc Natl Acad Sci U S A. 1989 Jan;86(2):665–669. doi: 10.1073/pnas.86.2.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang S. H., Reddick R. L., Piedrahita J. A., Maeda N. Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science. 1992 Oct 16;258(5081):468–471. doi: 10.1126/science.1411543. [DOI] [PubMed] [Google Scholar]