INFLUENCE OF TRIACYLGLYCEROL BIOSYNTHESIS RATE ON THE ASSEMBLY OF APO-B-100-CONTAINING LIPOPROTEINS IN HEP G2 CELLS

被引:83
作者
BOREN, J
RUSTAEUS, S
WETTESTEN, M
ANDERSSON, M
WIKLUND, A
OLOFSSON, SO
机构
[1] UNIV GOTEBORG,DEPT MED BIOCHEM,S-41390 GOTHENBURG,SWEDEN
[2] UNIV GOTEBORG,WALLENBERG LAB,GOTHENBURG,SWEDEN
来源
ARTERIOSCLEROSIS AND THROMBOSIS | 1993年 / 13卷 / 12期
关键词
APO-B-100; HEP G2 CELLS; TRIACYLGLYCEROL BIOSYNTHESIS;
D O I
10.1161/01.ATV.13.12.1743
中图分类号
R5 [内科学];
学科分类号
1002 [临床医学]; 100201 [内科学];
摘要
Apolipoprotein B-100 (apoB-100) appears in three forms in the endoplasmic reticulum of Hep G2 cells: (1) tightly bound to the membrane, ie, not extractable by sodium carbonate. This form is glycosylated but protease sensitive when present in intact microsomes, suggesting that it is only partially translocated to the microsomal lumen; (2) extractable by sodium carbonate and present on low-density lipoprotein-very-low-density lipoprotein (LDL-VLDL)-like particles. This form is glycosylated and secreted into the medium; and (3) extractable by sodium carbonate but having a higher density than the LDL-VLDL-like particles. This form, referred to as Fraction I, is glycosylated and protected against proteases when present in intact microsomal vesicles, indicating that it is completely translocated to the luminal side of the microsomal membrane. Fraction I is not secreted into the medium, but it disappears with time from the cell, suggesting that it is degraded. Oleic acid induced a 2.7-fold increase in the rate of the biosynthesis of triacylglycerol but not of phosphatidylcholine in Hep G2 cells. Incubation of the cells with oleic acid had no significant effect on the rate of initiation of the apoB-100-containing lipoproteins, nor did it influence the amount of apoB-100 that was associated with the membrane or the turnover of apoB-100 in the membrane. Instead, it increased the proportion of the nascent apoB polypeptides on initiated lipoproteins that was converted into full-length apoB-100 on LDL-VLDL-like particles, giving rise to an increased amount of these particles in the lumen of the secretory pathway. Pulse-chase experiments showed that incubation with oleic acid gave rise to an increased formation of LDL-VLDL-like particles ion behalf of the formation of Fraction I. This effect of oleic acid could partially explain the protective effect of the fatty acid on apoB-100, preventing it from undergoing posttranslational degradation.
引用
收藏
页码:1743 / 1754
页数:12
相关论文
共 21 条
[1]
Boren J., Graham L., Wettesten M., Scott J., White A., Olofsson S.-O., The assembly and secretion of apoB 100 containing lipoproteins in Hep G2 cells: ApoB 100 is cotranslationally integrated into lipoproteins, J Biol Chem, 267, pp. 9858-9867, (1992)
[2]
Boren J., Wettesten M., Sjoberg A., Thorlin T., Bondjers G., Wiklund O., Olofsson S.-O., The assembly and secretion of apoB 100 containing lipoproteins in Hep G2 cells: Evidence for different sites for protein synthesis and lipoprotein assembly, J Biol Chem, 265, pp. 10556-10564, (1990)
[3]
Spring D.J., Chen-Liu L.W., Chatterton J.E., Elovson J., Schumaker V.N., Lipoprotein assembly: Apolipoprotein B size determines lipoprotein core circumference, J Biol Chem, 267, pp. 14839-14845, (1992)
[4]
Bostrom K., Boren J., Wettesten M., Sjoberg A., Bondjers G., Wiklund O., Carlsson P., Olofsson S.-O., Studies on the assembly of apo B-100-containing lipoproteins in Hep G2 cells, J Biol Chem, 263, pp. 4434-4442, (1988)
[5]
Moberly J.B., Cole T.G., Alpers D.H., Schonfeld G., Oleic acid stimulation of apolipoprotein B secretion from Hep G2 cells occurs post-transcriptionally, Biochim Biophys Acta, 1042, pp. 70-80, (1990)
[6]
Pullinger C.R., North J.D., Teng B.-B., Rifici V.A., Ronhild De Brito A.E., Scott J., The apolipoprotein B gene is constitutively expressed in Hep G2 cells: Regulation of secretion by oleic acid, albumin, and insulin, and measurement of the mRNA half-life, J Lipid Res, 30, pp. 1065-1077, (1989)
[7]
Dixon J.L., Furukawa S., Ginsberg H.N., Olcate stimulates secretion of apolipoprotein B-containing lipoproteins from Hep G2 cells by inhibiting early intracellular degradation of apolipoprotein B, J Biol Chem, 266, pp. 5080-5086, (1991)
[8]
Yao Z., Vance D.E., Head group specificity in the requirement of phosphatidylcholine biosynthesis for very low density lipoprotein secretion from cultured hepatocytes, J Biol Chem, 264, pp. 11373-11380, (1989)
[9]
Yao Z., Vance D., The active synthesis of phosphatidylcholine is required for very low density lipoprotein secretion from rat hepatocytes, J Biol Chem, 263, pp. 2998-3004, (1988)
[10]
Wettesten M., Bostrom K., Bondjers G., Jarfeldt M., Norfeldt P.-I., Carella M., Wiklund O., Boren J., Olofsson S.-O., Pulse-chase studies of the synthesis of apolipoprotein B in a human hepatoma cell line, Hep G2, Eur J Biochem, 149, pp. 461-466, (1985)