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人类载脂蛋白B的合成:肝脏仅合成载脂蛋白B-100。

Apolipoprotein B synthesis in humans: liver synthesizes only apolipoprotein B-100.

作者信息

Edge S B, Hoeg J M, Schneider P D, Brewer H B

出版信息

Metabolism. 1985 Aug;34(8):726-30. doi: 10.1016/0026-0495(85)90022-8.

Abstract

Apolipoprotein (apo) B-100 and B-48 are prominent apolipoproteins in VLDL, IDL, and chylomicrons. Organ cultures of normal adult human liver were established to ascertain the form of apo B synthesized by hepatocytes in humans. Human liver was minced and incubated in 15 mL methionine-free RPMI-1640 medium with 10% dialyzed fetal calf serum plus 250 microCi 35S-methionine for eight hours at 37 degrees C. Lipoproteins secreted by the liver were isolated by ultracentrifugation and the content of newly synthesized apo B determined by quantitation of radioactivity in the apoB-100 and apoB-48 bands after separation by 3% NaDodSO4 gel electrophoresis. In the eight-hour period, 2.5% to 3.2% of added 35S-methionine was secreted in TCA-precipitable protein of which 0.34% was apo B. Ninety-nine percent of the apo B in VLDL, IDL, and LDL was in the apo B-100 electrophoretic band. No significant radioactivity was detected in the apo B-48 electrophoretic band. Eighty-nine percent of the total radioactivity of apo B-100 was in VLDL with 3% and 8% in IDL and LDL, respectively. These results establish that adult human liver in organ culture synthesizes apo B-100 but not apo B-48.

摘要

载脂蛋白(apo)B - 100和B - 48是极低密度脂蛋白(VLDL)、中间密度脂蛋白(IDL)和乳糜微粒中的主要载脂蛋白。建立正常成人肝脏的器官培养物以确定人类肝细胞合成的载脂蛋白B的形式。将人肝脏切碎,在含有10%透析胎牛血清加250微居里35S - 蛋氨酸的15毫升无蛋氨酸RPMI - 1640培养基中于37℃孵育8小时。通过超速离心分离肝脏分泌的脂蛋白,并通过在3%十二烷基硫酸钠(NaDodSO4)凝胶电泳分离后对apoB - 100和apoB - 48条带中的放射性进行定量来确定新合成的apo B的含量。在8小时内,添加的35S - 蛋氨酸中有2.5%至3.2%分泌到三氯乙酸(TCA)可沉淀蛋白中,其中0.34%是apo B。VLDL、IDL和低密度脂蛋白(LDL)中99%的apo B在apo B - 100电泳条带中。在apo B - 48电泳条带中未检测到明显放射性。apo B - 100总放射性的89%在VLDL中,分别有3%和8%在IDL和LDL中。这些结果表明,器官培养中的成人肝脏合成apo B - 100而非apo B - 48。

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