Biosynthesis and maturation of lactase-phlorizin hydrolase in the human small intestinal epithelial cells.
The biosynthesis and maturation of the human intestinal lactase-phlorizin hydrolase (LPH; EC 3.2.1.23-3.2.1.62) has been studied in cultured intestinal biopsies and mucosal explants. Short time pulse labelling revealed on high mannose intermediate of Mr 215,000 which was converted upon endo-beta-N-acetylglucosaminidase H (endo-H) digestion to a polypeptide of Mr 200,000. The brush border form of LPH was revealed after longer pulse periods and has Mr 160,000. It possesses mainly complex oligosaccharide chains and, owing to its partial endo-H sensitivity, at least one chain of the high mannose type. Leupeptin partially inhibited the appearance of the Mr-160,000 polypeptide. Monensin treatment of biopsies resulted in the modification of the Mr-160,000 species to the Mr-140,000 molecule, which was endo-H sensitive. Pulse-chase analysis indicated a slow post-translational processing of the high mannose precursor (Mr 215,000) to yield the mature brush-border form (Mr 160,000) of LPH. Our results further indicate that LPH is synthesized as a single polypeptide precursor which is intracellularly cleaved to yield the mature brush border of LPH. The data presented suggest that this cleavage occurs during the translocation of the molecule across the Golgi complex.
D000911:Antibodies, Monoclonal; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000378:metabolism; D004848:Epithelium / Q000201:enzymology; D005696:Galactosidases / Q000378:metabolism*; D005959:Glucosidases / Q000378:metabolism*; D006801:Humans; D007421:Intestine, Small / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism; D010694:Lactase-Phlorizin Hydrolase / Q000378:metabolism*; D007976:Leupeptins / Q000494:pharmacology; D008985:Monensin / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000378:metabolism*; D014176:Protein Biosynthesis; D014415:Tunicamycin / Q000494:pharmacology; D001616:beta-Galactosidase / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004792:Enzyme Precursors; D007527:Isoenzymes; D007976:Leupeptins; D009097:Multienzyme Complexes; D014415:Tunicamycin; D008985:Monensin; D005696:Galactosidases; D005959:Glucosidases; D001616:beta-Galactosidase; D010694:Lactase-Phlorizin Hydrolase; C032854:leupeptin