Abstract
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.
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- Andersson L. C., Gahmberg C. G. Surface glycoproteins of human white blood cells. Analysis by surface labeling. Blood. 1978 Jul;52(1):57–67. [PubMed] [Google Scholar]
- Antonowicz I., Lebenthal E. Developmental pattern of small intestinal enterokinase and disaccharidase activities in the human fetus. Gastroenterology. 1977 Jun;72(6):1299–1303. [PubMed] [Google Scholar]
- Barouki R., Finidori J., Chobert M. N., Aggerbeck M., Laperche Y., Hanoune J. Biosynthesis and processing of gamma-glutamyl transpeptidase in hepatoma tissue culture cells. J Biol Chem. 1984 Jun 25;259(12):7970–7974. [PubMed] [Google Scholar]
- Birkenmeier E., Alpers D. H. Enzymatic properties of rat lactase-phlorizin hydrolase. Biochim Biophys Acta. 1974 May 20;350(1):100–112. doi: 10.1016/0005-2744(74)90207-1. [DOI] [PubMed] [Google Scholar]
- Browning T. H., Trier J. S. Organ culture of mucosal biopsies of human small intestine. J Clin Invest. 1969 Aug;48(8):1423–1432. doi: 10.1172/JCI106108. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chow V., Murray R. K., Dixon J. D., Kurosky A. Biosynthesis of rabbit haptoglobin: chemical evidence for a single chain precursor. FEBS Lett. 1983 Mar 21;153(2):275–279. doi: 10.1016/0014-5793(83)80623-1. [DOI] [PubMed] [Google Scholar]
- DOELL R. G., KRETCHMER N. Studies of small intestine during development. I. Distribution and activity of beta-galactosidase. Biochim Biophys Acta. 1962 Aug 13;62:353–362. doi: 10.1016/0006-3002(62)90097-5. [DOI] [PubMed] [Google Scholar]
- Danielsen E. M., Skovbjerg H., Norén O., Sjöström H. Biosynthesis of intestinal microvillar proteins. Intracellular processing of lactase-phlorizin hydrolase. Biochem Biophys Res Commun. 1984 Jul 18;122(1):82–90. doi: 10.1016/0006-291x(84)90442-x. [DOI] [PubMed] [Google Scholar]
- Danielsen E. M., Skovbjerg H., Norén O., Sjöström H. Biosynthesis of intestinal microvillar proteins. Nature of precursor forms of microvillar enzymes from Ca2+-precipitated enterocyte membranes. FEBS Lett. 1981 Sep 28;132(2):197–200. doi: 10.1016/0014-5793(81)81159-3. [DOI] [PubMed] [Google Scholar]
- Docherty K., Steiner D. F. Post-translational proteolysis in polypeptide hormone biosynthesis. Annu Rev Physiol. 1982;44:625–638. doi: 10.1146/annurev.ph.44.030182.003205. [DOI] [PubMed] [Google Scholar]
- Elder J. H., Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. doi: 10.1073/pnas.79.15.4540. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fitting T., Kabat D. Evidence for a glycoprotein "signal" involved in transport between subcellular organelles. Two membrane glycoproteins encoded by murine leukemia virus reach the cell surface at different rates. J Biol Chem. 1982 Dec 10;257(23):14011–14017. [PubMed] [Google Scholar]
- Freiburghaus A. U., Schmitz J., Schindler M., Rotthauwe H. W., Kuitunen P., Launiala K., Hadorn B. Protein patterns of brush-border fragments in congenital lactose malabsorption and in specific hypolactasia of the adult. N Engl J Med. 1976 May 6;294(19):1030–1032. doi: 10.1056/NEJM197605062941903. [DOI] [PubMed] [Google Scholar]
- Garoff H., Kondor-Koch C., Riedel H. Structure and assembly of alphaviruses. Curr Top Microbiol Immunol. 1982;99:1–50. doi: 10.1007/978-3-642-68528-6_1. [DOI] [PubMed] [Google Scholar]
- Hanley J. M., Haugen T. H., Heath E. C. Biosynthesis and processing of rat haptoglobin. J Biol Chem. 1983 Jun 25;258(12):7858–7869. [PubMed] [Google Scholar]
- Hauri H. P., Quaroni A., Isselbacher K. J. Biogenesis of intestinal plasma membrane: posttranslational route and cleavage of sucrase-isomaltase. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5183–5186. doi: 10.1073/pnas.76.10.5183. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hauri H. P., Sterchi E. E., Bienz D., Fransen J. A., Marxer A. Expression and intracellular transport of microvillus membrane hydrolases in human intestinal epithelial cells. J Cell Biol. 1985 Sep;101(3):838–851. doi: 10.1083/jcb.101.3.838. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hedo J. A., Kahn C. R., Hayashi M., Yamada K. M., Kasuga M. Biosynthesis and glycosylation of the insulin receptor. Evidence for a single polypeptide precursor of the two major subunits. J Biol Chem. 1983 Aug 25;258(16):10020–10026. [PubMed] [Google Scholar]
- Kenny A. J., Maroux S. Topology of microvillar membrance hydrolases of kidney and intestine. Physiol Rev. 1982 Jan;62(1):91–128. doi: 10.1152/physrev.1982.62.1.91. [DOI] [PubMed] [Google Scholar]
- Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Ledford B. E., Davis D. F. Kinetics of serum protein secretion by cultured hepatoma cells. Evidence for multiple secretory pathways. J Biol Chem. 1983 Mar 10;258(5):3304–3308. [PubMed] [Google Scholar]
- Lodish H. F., Kong N., Snider M., Strous G. J. Hepatoma secretory proteins migrate from rough endoplasmic reticulum to Golgi at characteristic rates. Nature. 1983 Jul 7;304(5921):80–83. doi: 10.1038/304080a0. [DOI] [PubMed] [Google Scholar]
- Mahoney W. C., Duksin D. Biological activities of the two major components of tunicamycin. J Biol Chem. 1979 Jul 25;254(14):6572–6576. [PubMed] [Google Scholar]
- Olden K., Pratt R. M., Yamada K. M. Role of carbohydrates in protein secretion and turnover: effects of tunicamycin on the major cell surface glycoprotein of chick embryo fibroblasts. Cell. 1978 Mar;13(3):461–473. doi: 10.1016/0092-8674(78)90320-3. [DOI] [PubMed] [Google Scholar]
- Owen M. J., Kissonerghis A. M., Lodish H. F., Crumpton M. J. Biosynthesis and maturation of HLA-DR antigens in vivo. J Biol Chem. 1981 Sep 10;256(17):8987–8993. [PubMed] [Google Scholar]
- RUBINO A., ZIMBALATTI F., AURICCHIO S. INTESTINAL DISACCHARIDASE ACTIVITIES IN ADULT AND SUCKLING RATS. Biochim Biophys Acta. 1964 Nov 22;92:305–311. doi: 10.1016/0926-6569(64)90187-7. [DOI] [PubMed] [Google Scholar]
- Sahi T. Dietary lactose and the aetiology of human small-intestinal hypolactasia. Gut. 1978 Nov;19(11):1074–1086. doi: 10.1136/gut.19.11.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlegel-Haueter S., Hore P., Kerry K. R., Semenza G. The preparation of lactase and glucoamylase of rat small intestine. Biochim Biophys Acta. 1972 Feb 28;258(2):506–519. doi: 10.1016/0005-2744(72)90242-2. [DOI] [PubMed] [Google Scholar]
- Schmitz J., Preiser H., Maestracci D., Ghosh B. K., Cerda J. J., Crane R. K. Purification of the human intestinal brush border membrane. Biochim Biophys Acta. 1973 Sep 27;323(1):98–112. doi: 10.1016/0005-2736(73)90434-3. [DOI] [PubMed] [Google Scholar]
- Sjöström H., Norén O., Christiansen L., Wacker H., Semenza G. A fully active, two-active-site, single-chain sucrase.isomaltase from pig small intestine. Implications for the biosynthesis of a mammalian integral stalked membrane protein. J Biol Chem. 1980 Dec 10;255(23):11332–11338. [PubMed] [Google Scholar]
- Sjöström H., Norén O., Danielsen E. M., Skovbjerg H. Structure of microvillar enzymes in different phases of their life cycles. Ciba Found Symp. 1983;95:50–72. doi: 10.1002/9780470720769.ch5. [DOI] [PubMed] [Google Scholar]
- Strous G. J., Lodish H. F. Intracellular transport of secretory and membrane proteins in hepatoma cells infected by vesicular stomatitis virus. Cell. 1980 Dec;22(3):709–717. doi: 10.1016/0092-8674(80)90547-4. [DOI] [PubMed] [Google Scholar]
- Tarentino A. L., Maley F. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus. J Biol Chem. 1974 Feb 10;249(3):811–817. [PubMed] [Google Scholar]
- Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
- Umezawa H. Structures and activities of protease inhibitors of microbial origin. Methods Enzymol. 1976;45:678–695. doi: 10.1016/s0076-6879(76)45058-9. [DOI] [PubMed] [Google Scholar]
- Welsh J. D., Poley J. R., Bhatia M., Stevenson D. E. Intestinal disaccharidase activities in relation to age, race, and mucosal damage. Gastroenterology. 1978 Nov;75(5):847–855. [PubMed] [Google Scholar]
- Yamada K. M., Olden K. Fibronectins--adhesive glycoproteins of cell surface and blood. Nature. 1978 Sep 21;275(5677):179–184. doi: 10.1038/275179a0. [DOI] [PubMed] [Google Scholar]