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PMID: 7657695 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Hierarchy of mechanisms involved in generating Na/K-ATPase polarity in MDCK epithelial cells.

The Journal of cell biology ·Vol. 130 ·No. 5 ·1995-09-00 ·Pages 1105-15

Mays RW, Siemers KA, Fritz BA, Lowe AW, van Meer G, Nelson WJ

Abstract

We have studied mechanisms involved in generating a polarized distribution of Na/K-ATPase in the basal-lateral membrane of two clones of MDCK II cells. Both clones exhibit polarized distributions of marker proteins of the apical and basal-lateral membranes, including Na/K-ATPase, at steady state. Newly synthesized Na/K-ATPase, however, is delivered from the Golgi complex to both apical and basal-lateral membranes of one clone (II/J), and to the basal-lateral membrane of the other clone (II/G); Na/K-ATPase is selectively retained in the basal-lateral membrane resulting in the generation of complete cell surface polarity in both clones. Another basal-lateral membrane protein, E-cadherin, is sorted to the basal-lateral membrane in both MDCK clones, demonstrating that there is not a general sorting defect for basal-lateral membrane proteins in clone II/J cells. A glycosyl-phosphatidylinositol (GPI)-anchored protein (GP-2) and a glycosphingolipid (glucosylceramide, GlcCer) are preferentially transported to the apical membrane in clone II/G cells, but, in clone II/J cells, GP-2 and GlcCer are delivered equally to both apical and basal-lateral membranes, similar to Na/K-ATPase. To examine this apparent inter-relationship between sorting of GlcCer, GP-2 and Na/K-ATPase, sphingolipid synthesis was inhibited in clone II/G cells with the fungal metabolite, Fumonisin B1 (FB1). In the presence of FB1, GP-2 and Na/K-ATPase are delivered to both apical and basal-lateral membranes, similar to clone II/J cells; FB1 had no effect on sorting of E-cadherin to the basal-lateral membrane of II/G cells. Addition of exogenous ceramide, to circumvent the FB1 block, restored GP-2 and Na/K-ATPase sorting to the apical and basal-lateral membranes, respectively. These results show that the generation of complete cell surface polarity of Na/K-ATPase involves a hierarchy of sorting mechanisms in the Golgi complex and plasma membrane, and that Na/K-ATPase sorting in the Golgi complex of MDCK cells may be regulated by exclusion from an apical pathway(s). These results also provide new insights into sorting pathways for other apical and basal-lateral membrane proteins.

MeSH Terms
Animals Cadherins/metabolism Cell Polarity/physiology Cells, Cultured Dogs Epithelial Cells Fumonisins Glycosphingolipids/antagonists & inhibitors,biosynthesis,metabolism Glycosylphosphatidylinositols/metabolism Golgi Apparatus/metabolism Kidney/cytology Membrane Glycoproteins/metabolism Membrane Proteins/metabolism Mycotoxins/pharmacology Protein Biosynthesis Proteins/metabolism Rabbits Rats Sodium-Potassium-Exchanging ATPase/metabolism Teratogens/pharmacology
Chemicals
Cadherins Fumonisins Glycosphingolipids Glycosylphosphatidylinositols Membrane Glycoproteins Membrane Proteins Mycotoxins Proteins Teratogens glycoprotein-2, pancreas fumonisin B1 Sodium-Potassium-Exchanging ATPase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Mays R W
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, California 94305, USA.
Siemers K A
Fritz B A
Lowe A W
van Meer G
Nelson W J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.130.5.1105
Published
1995-09-00
Pages
1105-15
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120560
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.130.5.1105 PMC Full Text View journal details
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