Apolipoprotein B synthesis by human liver and intestine in vitro

Abstract

The synthesis of apolipoprotein B (apoB) was examined in human fetal and adult intestine and liver. Intestine and liver were minced and then incubated with [3H]leucine, homogenized, and subjected to immunoprecipitation with antiserum that recognized both apoB-100 and apoB-48 (forms of apoB found in low density lipoproteins and in chylomicrons, respectively). Immunoprecipitates of fetal and adult liver contained radioactivity in a single apoB-100 peak when examined by NaDodSO4/polyacrylamide gel electrophoresis. Intestine from fetuses at 11 weeks of gestation incorporated radioactivity mainly into apoB-100, with little incorporation into apoB-48. Sixteen-week fetal intestine showed both apoB-100 and apoB-48, whereas adult intestine incorporated radioactivity only into apoB-48. Pulse-chase experiments with 11- and 16-week fetal intestine showed no evidence for the conversion of apoB-100 to apoB-48. Incubation of intestinal homogenates with fetal liver apoB-100 did not result in the conversion of apoB-100 to smaller forms of apoB. A cDNA probe to hepatic apoB-100 identified a single, 18-kilobase transcript in poly(A)+ RNA from fetal and adult liver and fetal intestine of all ages. These studies define the developmental pattern of apoB synthesis in human fetal and adult liver and intestine. No evidence could be found for the conversion of apoB-100 to apoB-48. The finding of a single mRNA transcript despite the form of apoB synthesized in each tissue is discussed.