Biosynthetic precursor (214 kDa) of apolipoprotein B-48 is not secreted by Caco-2 cells and normal human intestine.
The synthesis and secretion of apolipoprotein B (apo B) was studied in a human colon carcinoma (Caco-2) cell line and in explants from normal human intestine. In Caco-2 cells, the specific activity of the intestinal disaccharidases maltase, sucrase-isomaltase and lactase was enhanced 8-, 6- and 3-fold respectively, at 19 days post-confluence as compared with 1-day-post-confluence cultures. The level of apo B secreted into the medium increased from undetectable in the cells just reaching confluency, to 115 ng/ml at 18 days post-confluence. The presence of apo B-100 and apo B-48 with mobilities on SDS/polyacrylamide-gel electrophoresis corresponding to those of human very-low-density lipoproteins and lymph chylomicrons, respectively, was detected in the media from 7-, 12- and 18-days-post-confluence cells. These two apo B proteins were also found intracellularly in 7-day-post-confluence cultures. However, more differentiated cells (12 and 18 days post-confluence) accumulated large amount of a 214 kDa protein intracellularly. Apo B-related 214 kDa protein was also synthesized by normal human intestinal explants. A pulse-chase experiment with explants from normal human jejunum showed a slow intracellular conversion of the 214 kDa protein into the size of mature apo B-48 (264 kDa), concomitant with increasing amounts of mature apo B-48 in the medium, suggesting a precursor-product relationship. Despite large intracellular quantities, the 214 kDa protein from the normal human tissue and Caco-2 cells was absent from the medium. No apo B-100 synthesis was detected in the human explants. These findings may help in our understanding of cholesterol and lipid metabolism in health and in some disorders characterized by the inability to secrete apo B-containing lipoproteins.
Sasak
W V
WV
Department of Pediatrics, New England Medical Center, Tufts University School of Medicine, Boston, MA 02111.
Buller
H A
HA
Reinhold
R
R
D053283:Apolipoprotein B-48; D001055:Apolipoproteins B / Q000096:biosynthesis* / Q000302:isolation & purification; D002460:Cell Line; D003110:Colonic Neoplasms; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007421:Intestine, Small / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009924:Organ Culture Techniques; D011498:Protein Precursors / Q000096:biosynthesis* / Q000302:isolation & purification; D012016:Reference Values; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D053283:Apolipoprotein B-48; D001055:Apolipoproteins B; D011498:Protein Precursors