Home LiteratureArticle Details
PMID: 2687880 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 23 ·1989-12-00 ·Pages 9313-7

Le Bivic A, Real FX, Rodriguez-Boulan E

Abstract

We studied the surface delivery pathways followed by newly synthesized plasma membrane proteins in intestinal cells. To this end, we developed an assay and characterized an epithelial cell line (SK-CO-15) derived from human colon adenocarcinoma. Polarized confluent monolayers (2000 omega.cm2), grown on polycarbonate filter chambers, were pulsed with radioactive methionine/cysteine and, at different times of chase, the protein fraction reaching the apical or basolateral surface was recovered by domain-selective biotinylation, immunoprecipitation, and immobilized streptavidin precipitation. Both an apical and a basolateral marker were found to be delivered vectorially to the respective surface, with a sorting efficiency of 50:1 for the basolateral marker and 14:1 for the apical marker.

MeSH Terms
Adenocarcinoma Cell Line Cell Membrane/metabolism,ultrastructure Colonic Neoplasms Epithelium/metabolism Fluorescent Antibody Technique Humans Kinetics Membrane Proteins/biosynthesis Microscopy, Electron Neoplasm Proteins/biosynthesis Tumor Cells, Cultured/metabolism,ultrastructure
Chemicals
Membrane Proteins Neoplasm Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Le Bivic A
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.
Real F X
Rodriguez-Boulan E
References (38)
38 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. A technique for ultracryotomy of cell suspensions and tissues.
    J Cell Biol. 1973 May;57(2):551-65 PMID: 4121290
  3. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus.
    J Biol Chem. 1974 Feb 10;249(3):811-7 PMID: 4204552
  4. Transepithelial transport in cell culture.
    Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212-6 PMID: 1063404
  5. Synthesis of membrane glycoproteins in rat small-intestinal villus cells. Redistribution of L-[1,5,6-3H]fucose-labelled membrane glycoproteins among Golgi, lateral basal and microvillus membranes in vivo.
    Biochem J. 1979 Jul 15;182(1):203-12 PMID: 496908
  6. Biogenesis of intestinal plasma membrane: posttranslational route and cleavage of sucrase-isomaltase.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):5183-6 PMID: 291933
  7. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate.
    Anal Biochem. 1979 Sep 15;98(1):132-5 PMID: 543547
  8. Phase separation of integral membrane proteins in Triton X-114 solution.
    J Biol Chem. 1981 Feb 25;256(4):1604-7 PMID: 6257680
  9. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  10. Localization by immunofluorescence and histochemical labeling of aminopeptidase N in relation to its biosynthesis in rabbit and pig enterocytes.
    Gastroenterology. 1982 Feb;82(2):317-24 PMID: 6172312
  11. 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):4540-4 PMID: 6812050
  12. Use of immunocytochemical techniques in studying the biogenesis of cell surfaces in polarized epithelia.
    Methods Enzymol. 1983;98:379-95 PMID: 6366469
  13. Polarized assembly of enveloped viruses from cultured epithelial cells.
    Methods Enzymol. 1983;98:486-501 PMID: 6321907
  14. Sorting of an apical plasma membrane glycoprotein occurs before it reaches the cell surface in cultured epithelial cells.
    J Cell Biol. 1984 Dec;99(6):2131-9 PMID: 6501415
  15. Biogenesis of epithelial cell polarity: intracellular sorting and vectorial exocytosis of an apical plasma membrane glycoprotein.
    Cell. 1984 Dec;39(3 Pt 2):537-46 PMID: 6509551
  16. Polarized delivery of viral glycoproteins to the apical and basolateral plasma membranes of Madin-Darby canine kidney cells infected with temperature-sensitive viruses.
    J Cell Biol. 1985 Jan;100(1):136-51 PMID: 2981229
  17. Intracellular sorting and basolateral appearance of the G protein of vesicular stomatitis virus in Madin-Darby canine kidney cells.
    J Cell Biol. 1985 Aug;101(2):470-6 PMID: 2991300
  18. Immuno-electronmicroscopical localization of a microvillus membrane disaccharidase in the human small-intestinal epithelium with monoclonal antibodies.
    Eur J Cell Biol. 1985 Jul;38(1):6-15 PMID: 3896809
  19. Biosynthesis of intestinal microvillar proteins. Evidence for an intracellular sorting taking place in, or shortly after, exit from the Golgi complex.
    Eur J Biochem. 1985 Oct 15;152(2):493-9 PMID: 2865138
  20. Polymeric immunoglobulin receptor expressed in MDCK cells transcytoses IgA.
    Cell. 1986 Aug 15;46(4):613-21 PMID: 3524859
  21. Intracellular sorting and polarized cell surface delivery of (Na+,K+)ATPase, an endogenous component of MDCK cell basolateral plasma membranes.
    Cell. 1986 Aug 15;46(4):623-31 PMID: 3015421
  22. Localization of sucrase-isomaltase in the rat enterocyte.
    Gastroenterology. 1987 Jan;92(1):98-105 PMID: 3536655
  23. The sorting of proteins to the plasma membrane in epithelial cells.
    J Cell Biol. 1986 Dec;103(6 Pt 2):2565-8 PMID: 3025218
  24. Cell surface polarity in epithelia.
    Annu Rev Cell Biol. 1985;1:243-88 PMID: 3939606
  25. Evidence for the transit of aminopeptidase N through the basolateral membrane before it reaches the brush border of enterocytes.
    J Membr Biol. 1987;96(1):19-25 PMID: 2884323
  26. Changes in villin synthesis and subcellular distribution during intestinal differentiation of HT29-18 clones.
    J Cell Biol. 1987 Jul;105(1):359-69 PMID: 2440895
  27. Biogenesis of the rat hepatocyte plasma membrane in vivo: comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation.
    J Cell Biol. 1987 Sep;105(3):1241-51 PMID: 3654750
  28. Apical membrane marker is expressed early in colonic epithelial cells.
    Biol Cell. 1987;60(3):209-16 PMID: 2962680
  29. HT-29 cells are an in vitro model for the generation of cell polarity in epithelia during embryonic differentiation.
    Proc Natl Acad Sci U S A. 1988 Jan;85(1):136-40 PMID: 3277169
  30. Dissection of the asynchronous transport of intestinal microvillar hydrolases to the cell surface.
    J Cell Biol. 1988 Jun;106(6):1853-61 PMID: 2898478
  31. Characterization of a glycoprotein expressed on the basolateral membrane of human intestinal epithelial cells and cultured colonic cell lines.
    Eur J Cell Biol. 1988 Apr;46(1):113-20 PMID: 3294003
  32. Polarized apical distribution of glycosyl-phosphatidylinositol-anchored proteins in a renal epithelial cell line.
    Proc Natl Acad Sci U S A. 1988 Dec;85(24):9557-61 PMID: 2974157
  33. Nocodazole, a microtubule-active drug, interferes with apical protein delivery in cultured intestinal epithelial cells (Caco-2).
    J Cell Biol. 1989 Jan;108(1):13-22 PMID: 2642910
  34. A new method for detecting endocytosed proteins.
    EMBO J. 1988 Dec 20;7(13):4087-92 PMID: 3243274
  35. External and internal signals for epithelial cell surface polarization.
    Annu Rev Physiol. 1989;51:741-54 PMID: 2653203
  36. Integral and peripheral protein composition of the apical and basolateral membrane domains in MDCK cells.
    J Membr Biol. 1989 Mar;107(3):277-86 PMID: 2716048
  37. A sorting signal for the basolateral delivery of the vesicular stomatitis virus (VSV) G protein lies in its luminal domain: analysis of the targeting of VSV G-influenza hemagglutinin chimeras.
    Proc Natl Acad Sci U S A. 1989 Jun;86(11):4112-6 PMID: 2542964
  38. Plasma membrane protein sorting in epithelial cells: do secretory pathways hold the key?
    Trends Biochem Sci. 1988 May;13(5):181-4 PMID: 3255200
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.23.9313
Published
1989-12-00
Pages
9313-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298485
Subset
IM
Grants
NIGMS NIH HHS · R01-GM34107 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.23.9313 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2