[en] We have identified a G-to-A transition in exon 3 of the APOC3 gene resulting in a novel Ala23Thr apolipoprotein (apo) C-III variant, associated with apoC-III deficiency in three unrelated Yucatan Indians. The Ala23Thr substitution modifies the hydrophobic/hydrophilic repartition of the helical N-terminal peptide and hence could disturb the lipid association. In vitro expression in Escherichia coli of wild-type and mutant apoC-III enabled the characterization of the variant. Compared with wild-type apoC-III-Ala23, the mutant apoC-III-Thr23 showed reduced affinity for dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles with higher amounts of free apoC-III. Displacement of apoE from discoidal apoE:dipalmitoylphosphatidycholine (DPPC) complex by apoC-III-Thr23 was comparable to wild type but the less efficient binding of the apoC-III-Thr23 to the discoidal complex resulted in a higher apoE/apoC-III (mol/mol) ratio (34%) than with wild-type/apoE:DPPC mixtures. The inhibition of lipoprotein lipase (LPL) by apoC-III-Thr23 was comparable to that of wild type, and therefore effects on LPL activity could not explain the lower triglyceride (Tg) levels in Thr-23 carriers. Thus, these in vitro results suggest that in vivo the less efficient lipid binding of apoC-III-Thr23 might lead to a faster catabolism of free apoC-III, reflected in the reduced plasma apoC-III levels identified in Thr-23 carriers, and poorer competition with apoE, which might enhance clearance of Tg-rich lipoproteins and lower plasma Tg levels seen in Thr-23 carriers.
Herbert P.N., Assmann G., Grotton J.A.M., Fredrickson D.S. (1999) Disorders of lipoprotein and lipid metabolism., The Metabolic Basis of Inherited Disease. J. B. Stanbury, D. S. Wyngaarden, D. S. Fredrickson, J. L. Goldstein, and M. S. Brown, editors. McGraw-Hill, New York; 589-651.
Shoulders C.C., Harry P.J., Lagrost L., White S.E., Shah N.F., North J.D., Gilligan M., Gambert P., Ball M.J. (1991) Variation at the apo AI/CIII/AIV gene complex is associated with elevated plasma levels of apo CIII. Atherosclerosis 87:239-247.
Le N.A., Gibson J.C., Ginsberg H.N. (1988) Independent regulation of plasma apolipoprotein C-II and C-III concentrations in very low density and high density lipoproteins: Implications for the regulation of the catabolism of these lipoproteins. J. Lipid Res. 29:669-677.
Marz W., Schenk G., Gross W. (1987) Apolipoproteins C-II and C-III in serum quantified by zone immunoelectrophoresis. Clin. Chem. 33:664-669.
Chivot L., Mainard F., Bigot E., Bard J.M., Auget J.L., Madec Y., Fruchart J.C. (1990) Logistic discriminant analysis of lipids and apolipoproteins in a population of coronary bypass patients and the significance of apolipoproteins C-III and E. Atherosclerosis 82:205-211.
Wiseman S.A., Powell J.T., Barber N., Humphries S.E., Greenhalgh R.M. (1991) Influence of apolipoproteins on the anatomical distribution of arterial disease. Atherosclerosis 89:231-237.
Blankenhorn D.H., Selzer R.H., Crawford D.W., Barth J.D., Liu C.R., Liu C.H., Mack W.J., Alaupovic P. (1993) Beneficial effects of colestipol-niacin therapy on the common carotid artery. Two- and four-year reduction of intima-media thickness measured by ultrasound. Circulation 88:20-28.
Hodis H.N., Mack W.J., Azen S.P., Alaupovic P., Pogoda J.M., LaBree L., Hemphill L.C., Kramsch D.M., Blankenhorn D.H. (1994) Triglyceride- and cholesterol-rich lipoproteins have a differential effect on mild/moderate and severe lesion progression as assessed by quantitative coronary angiography in a controlled trial of lovastatin. Circulation 90:42-49.
Brown W.V., Baginsky M.L. (1972) Inhibition of lipoprotein lipase by an apoprotein of human very low density lipoprotein. Biochem. Biophys. Res. Commun. 46:375-382.
Ginsberg H.N., Le N.A., Goldberg I.J., Gibson J.C., Rubinstein A., Wang Iverson P., Norum R., Brown W.V. (1986) Apolipoprotein B metabolism in subjects with deficiency of apolipoproteins CIII and AI. Evidence that apolipoprotein CIII inhibits catabolism of triglyceride-rich lipoproteins by lipoprotein lipase in vivo. J. Clin. Invest. 78:1287-1295.
Wang C.S., McConathy W.J., Kloer H.U., Alaupovic P. (1985) Modulation of lipoprotein lipase activity by apolipoproteins. Effect of apolipoprotein C-III. J. Clin. Invest. 75:384-390.
Quarfordt S.H., Michalopoulos G., Schirmer B. (1982) The effect of human C apolipoproteins on the in vitro hepatic metabolism of triglyceride emulsions in the rat. J. Biol. Chem. 257:14642-14647.
Windler E., Havel R.J. (1985) Inhibitory effects of C apolipoproteins from rats and humans on the uptake of triglyceride-rich lipoproteins and their remnants by the perfused rat liver. J. Lipid Res. 26:556-565.
Sehayek E., Eisenberg S. (1991) Mechanisms of inhibition by apolipoprotein C of apolipoprotein E-dependent cellular metabolism of human triglyceride-rich lipoproteins through the low density lipoprotein receptor pathway. J. Biol. Chem. 266:18259-18267.
Maeda N., Li H., Lee D., Oliver P., Quarfordt S.H., Osada J. (1994) Targeted disruption of the apolipoprotein C-III gene in mice results in hypotriglyceridemia and protection from postprandial hypertriglyceridemia. J. Biol. Chem. 269:23610-23616.
Ito Y., Azrolan N., O'Connell A., Walsh A., Breslow J.L. (1990) Hypertriglyceridemia as a result of human apo CIII gene expression in transgenic mice. Science 249:790-793.
Aalto Setala K., Kontula K., Sane T., Nieminen M., Nikkila E. (1987) DNA polymorphisms of apolipoprotein A-I/C-III and insulin genes in familial hypertriglyceridemia and coronary heart disease. Atherosclerosis 66:145-152.
Ebara T., Ramakrishnan R., Steiner G., Shachter N.S. (1997) Chylomicronemia due to apolipoprotein CIII overexpression in apolipoprotein E-null mice. Apolipoprotein CIII-induced hypertriglyceridemia is not mediated by effects on apolipoprotein. J. Clin. Invest. 99:2672-2681.
Karathanasis S.K., McPherson J., Zannis V.I., Breslow J.L. (1983) Linkage of human apolipoproteins A-I and C-III genes. Nature 304:371-373.
Ordovas J.M., Civeira F., Genest J. Jr., Craig S., Robbins A.H., Meade T., Pocovi M., Frossard P.M., Masharani U., Wilson P.W., Salem D.N., Ward R.H., Schaefer E.J. (1991) Restriction fragment length polymorphisms of the apolipoprotein A-I, C-III, A-IV gene locus. Relationships with lipids, apolipoproteins, and premature coronary artery disease. Atherosclerosis 87:75-86.
Xu C.F., Talmud P., Schuster H., Houlston R., Miller G., Humphries S. (1994) Association between genetic variation at the APO AI-CIII-AIV gene cluster and familial combined hyperlipidaemia. Clin. Genet. 46:385-397.
Rees A., Shoulders C.C., Stocks J., Galton D.J., Baralle F.E. (1983) DNA polymorphism adjacent to human apoprotein A-1 gene: Relation to hypertriglyceridaemia. Lancet 1:444-446.
Paulweber B., Friedl W., Krempler F., Humphries S.E., Sandhofer F. (1988) Genetic variation in the apolipoprotein AI-CIII-AIV gene cluster and coronary heart disease. Atherosclerosis 73:125-133.
Rees A., Stocks J., Sharpe C.R., Vella M.A., Shoulders C.C., Katz J., Jowett N.I., Baralle F.E., Galton D.J. (1985) Deoxyribonucleic acid polymorphism in the apolipoprotein A-1-C-III gene cluster. Association with hypertriglyceridemia. J. Clin. Invest. 76:1090-1095.
Ferns G.A., Stocks J., Ritchie C., Galton D.J. (1985) Genetic polymorphisms of apolipoprotein C-III and insulin in survivors of myocardial infarction. Lancet 2:300-303.
Karathanasis S.K., Ferris E., Haddad I.A. (1987) DNA inversion within the apolipoproteins AI/CIII/AIV-encoding gene cluster of certain patients with premature atherosclerosis. Proc. Natl. Acad. Sci. USA 84:7198-7202.
Ordovas J.M., Cassidy D.K., Civeira F., Bisgaier C.L., Schaefer E.J. (1989) Familial apolipoprotein A-I, C-III, and A-IV deficiency and premature atherosclerosis due to deletion of a gene complex on chromosome 11. J. Biol. Chem. 264:16339-16342.
Von Eckardstein A., Holz H., Sandkamp M., Weng W., Funke H., Assmann G. (1991) Apolipoprotein C-III(Lys58→Glu). Identification of an apolipoprotein C-III variant in a family with hyperalphalipoproteinemia. J. Clin. Invest. 87:1724-1731.
Maeda H., Hashimoto R.K., Ogura T., Hiraga S., Uzawa H. (1987) Molecular cloning of a human apoC-III variant: Thr 74→Ala 74 mutation prevents O-glycosylation. J. Lipid Res. 28:1405-1409.
Luttmann S., Von Eckardstein A., Wei W., Funke H., Kohler E., Mahley R.W., Assmann G. (1994) Electrophoretic screening for genetic variation in apolipoprotein C-III: Identification of a novel apoC-III variant, apoC-III(Asp45→Asn), in a Turkish patient. J. Lipid Res. 35:1431-1440.
Pullinger C.R., Malloy M.J., Shahidi A.K., Ghassemzadeh M., Duchateau P., Villagomez J., Allaart J., Kane J.P. (1997) A novel apolipoprotein C-III variant, apoC-III(Gln38→Lys), associated with moderate hypertriglyceridemia in a large kindred of Mexican origin. J. Lipid Res. 38:1833-1840.
Ferrell R.E., Kamboh M.I., Sepehrnia B.S., Adams Campbell L.L., Weiss K.M. (1988) Genetic variation in the apolipoproteins C-II and C-III. Adv. Exp. Med. Biol. 243:81-85.
Mancini G., Carbonara A.O., Heremans J.F. (1965) Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 2:235-254.
Allen C.C., Poon L.S., Chan C.S.G., Richmond M., Fu P.C. (1999) Enzymatic determination of total serum cholesterol. Clin. Chem. 20:470-475.
Ahn Y.I., Valdez R., Reddy A.P., Cole S.A., Weiss K.M., Ferrell R.E. (1991) DNA polymorphisms of the apolipoprotein AI/CIII/AIV gene cluster influence plasma cholesterol and triglyceride levels in the Mayans of the Yucatan Peninsula, Mexico. Hum. Hered. 41:281-289.
Brasseur R., Lins L., Vanloo B., Ruysschaert J.M., Rosseneu M. (1992) Molecular modeling of the amphipathic helices of the plasma apolipoproteins. Proteins 13:246-257.
Brasseur R. (1990) Theoretical analysis of molecular membrane organization., Molecular Description of Biological Membrane Components by Computer-Aided Conformational Analysis. R. Brasseur, editor. CRC Press, Boca Raton, FL; 1:203-219.
Brasseur R. (1991) Differentiation of lipid-associating helices by use of three-dimensional molecular hydrophobicity potential calculations. J. Biol. Chem. 266:16120-16127.
Geourjon C., Deleage G. (1995) SOPMA: Significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Comput. Appl. Biosci. 11:681-684.
Rost B., Sander C. (1993) Prediction of protein secondary structure at better than 70% accuracy. J. Mol. Biol. 232:584-599.
Rost B., Sander C. (1994) Combining evolutionary information and neural networks to predict protein secondary structure. Proteins 19:55-72.
Frishman D., Argos P. (1996) Incorporation of non-local interactions in protein secondary structure prediction from the amino acid sequence. Protein Eng 9:133-142.
Garnier J., Gibrat J.F., Robson B. (1996) GOR method for predicting protein secondary structure from amino acid sequence. Methods Enzymol. 266:540-553.
Deleage G., Roux B. (1987) An algorithm for protein secondary structure prediction based on class prediction. Protein Eng. 1:289-294.
King R.D., Sternberg M.J. (1996) Identification and application of the concepts important for accurate and reliable protein secondary structure prediction. Protein Sci. 5:2298-2310.
Levin J.M., Robson B., Garnier J. (1986) An algorithm for secondary structure determination in proteins based on sequence similarity. FEBS Lett 205:303-308.
Levin J.M. (1997) Exploring the limits of nearest neighbour secondary structure prediction. Protein Eng. 10:771-776.
Guermeur Y., Geourjon C., Gallinari P., Deleage G. (1999) Improved performance in protein secondary structure prediction by inhomogeneous score combination. Bioinformatics 15:413-421.
De Pauw M., Vanloo B., Weisgraber K., Rosseneu M. (1995) Comparison of lipid-binding and lecithin:cholesterol acyltransferase activation of the amino- and carboxyl-terminal domains of human apolipoprotein E3. Biochemistry 34:10953-10966.
Labeur C., Lambert G., Van Cauteren T., Duverger N., Vanloo B., Chambaz J., Vandekerckhove J., Castro G., Rosseneu M. (1998) Displacement of apo A-I from HDI, by apo A-II or its C-terminal helix promotes the formation of pre-betal migrating particles and decreases LCAT activation. Atherosclerosis 139:351-362.
Pillot T., Barbier A., Visvikis A., Lozac'h K., Rosseneu M., Vandekerckhove J., Siest G. (1996) Single-step purification of two functional human apolipoprotein E variants hyperexpressed in Escherichia coli. Protein Expr. Purif. 7:407-414.
Rosseneu M.Y., Labeur C. (1990) Apolipoprotein structure, function and measurement. Curr. Opin. Lipidol. 1:508-513.
Bury J., Rosseneu M. (1985) Quantification of human serum apolipoprotein AI by enzyme immunoassay. Clin. Chem. 31:247-251.
Bengtsson-Olivecrona G., Olivecrona T. (1992) Assay of lipoprotein lipase and hepatic lipase., Lipoprotein Analysis. A Practical Approach. C. A. Converse and E. R. Skinner, editors. Oxford Unversity Press, New York; 169-185.
Saheki S., Takahashi I., Murase M., Takeuchi N., Uchida K. (1991) Composition of very low density lipoproteins and in vitro effect of lipoprotein lipase. Clin. Chim. Acta 204:155-166.
Chisholm J.W., Gebre A.K., Parks J.S. (1999) Characterization of C-terminal histidine-tagged human recombinant lecithin:cholesterol acyltransferase. J. Lipid Res. 40:1512-1519.
Waterworth D.M., Ribalta J., Nicaud V., Dallongeville J., Humphries S.E., Talmud P. (1999) ApoC-III gene variants modulate postprandial response to both glucose and fat tolerance tests. Circulation 99:1872-1877.
Bengtsson Olivecrona G., Sletten K. (1990) Primary structure of the bovine analogues to human apolipoproteins CII and CIII. Studies on isoforms and evidence for proteolytic processing. Eur. J. Biochem. 192:515-521.
Li W.H., Tanimura M., Luo C.C., Datta S., Chan I. (1988) The apolipoprotein multigene family: Biosynthesis, structure, structure-function relationships, and evolution. J. Lipid Res. 29:245-271.
Luo C.C., Li W.H., Moore M.N., Chan L. (1986) Structure and evolution of the apolipoprotein multigene family. J. Mol. Biol. 187:325-340.
Datta S., Li W.H., Ghosh I., Luo C.C., Chan L. (1987) Structure and expression of dog apolipoprotein C-II and C-III mRNAs. Implications for the evolution and functional constraints of apolipoprotein structure. J. Biol. Chem. 262:10588-10593.
Jonas A. (1992) Lipid-binding properties of apolipoproteins., Structure and Function of Apolipoproteins. M. Rosseneu, editor. CRC Press, Ann Arbor, MI; 217-245.
Rogers D.P., Roberts L.M., Lebowitz J., Datta G., Anantharamaiah G.M., Engler J.A., Brouillette C.G. (1998) The lipid-free structure of apolipoprotein A-I: Effects of amino-terminal deletions. Biochemistry 37:11714-11725.
Bergeron J., Frank P.G., Emmanuel F., Latta M., Zhao Y., Sparks D.L., Rassart E., Denefle P., Marcel Y.L. (1997) Characterization of human apolipoprotein A-I expressed in Escherichia coli. Biochim. Biophys. Acta 1344:139-152.
Daum U., Langer C., Duverger N., Emmanuel F., Benoit P., Denefle P., Chirazi A., Cullen P., Pritchard P.H., Bruckert E., Assmann G., Von Eckardstein A. (1999) Apolipoprotein A-I (R151C) Paris is defective in activation of lecithin:cholesterol acyltransferase but not in initial lipid binding, formation of reconstituted lipoproteins, or promotion of cholesterol efflux. J. Mol. Med. 77:614-622.
Dong L.M., Innerarity T.L., Arnold K.S., Newhouse Y.M., Weisgraber K.H. (1998) The carboxyl terminus in apolipoprotein E2 and the seven amino acid repeat in apolipoprotein E-Leiden: Role in receptor-binding activity. J. Lipid Res. 39:1173-1180.
Aalto Setala K., Fisher F.A., Chen X., Chajek Shaul T., Hayek T., Zechner R., Walsh A., Ramakrishnan R., Ginsberg H.N., Breslow J.L. (1992) Mechanism of hypertriglyceridemia in human apolipoprotein (apo) CIII transgenic mice. Diminished very low density lipoprotein fractional catabolic rate associated with increased apo CIII and reduced apo E on the particles. J. Clin. Invest. 90:1889-1900.
Aalto Setala K., Weinstock P.H., Bisgaier C.L., Wu L., Smith J.D., Breslow J.L. (1996) Further characterization of the metabolic properties of triglyceride-rich lipoproteins from human and mouse apoC-III transgenic mice. J. Lipid Res. 37:1802-1811.
De Silva H.V., Lauer S.J., Wang J., Simonet W.S., Weisgraber K.H., Mahley R.W., Taylor J.M. (1994) Overexpression of human apolipoprotein C-III in transgenic mice results in an accumulation of apolipoprotein B48 remnants that is corrected by excess apolipoprotein E. J. Biol. Chem. 269:2324-2335.
Kowal R.C., Herz J., Weisgraber K.H., Mahley R.W., Brown M.S., Goldstein J.L. (1990) Opposing effects of apolipoproteins E and C on lipoprotein binding to low density lipoprotein receptor-related protein. J. Biol. Chem. 265:10771-10779.
Cardin A.D., Jackson R.L., Johnson J.D. (1982) 5-Dimethylaminonaphthalene-1-sulfonyl 3-aminotyrosyl apolipoprotein C-III. Preparation, characterization, and interaction with phospholipid vesicles. J. Biol. Chem. 257:4987-4992.
Dyer C.A., Smith R.S., Curtiss L.K. (1991) Only multimers of a synthetic peptide of human apolipoprotein E are biologically active. J. Biol. Chem. 266:15009-15015.