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PMID: 10982412 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Expression of podocalyxin inhibits cell-cell adhesion and modifies junctional properties in Madin-Darby canine kidney cells.

Molecular biology of the cell ·Vol. 11 ·No. 9 ·2000-09-00 ·Pages 3219-32

Takeda T, Go WY, Orlando RA, Farquhar MG

Abstract

Podocalyxin is a major membrane protein of the glomerular epithelium and is thought to be involved in maintenance of the architecture of the foot processes and filtration slits characteristic of this unique epithelium by virtue of its high negative charge. However, until now there has been no direct evidence for podocalyxin's function. Podocalyxin is a type 1 transmembrane sialoprotein with an N-terminal mucin-like domain. To assess its function, we cloned rat podocalyxin and examined the effects of its expression on the cell adhesion properties of stably transfected Chinese hamster ovary (CHO)-K1 and Madin-Darby canine kidney (MDCK) cells and inducible ecdysone receptor-expressing (EcR)-CHO cells. In a cell aggregation assay, CHO-K1 cells expressing high levels of podocalyxin showed complete inhibition of cell aggregation, and MDCK transfectants showed greatly reduced aggregation ( approximately 60-80%) compared with parental cells. In EcR-CHO cells, the expression level of podocalyxin induced by increasing levels of ecdysone analogue correlated closely with the antiadhesion effect. The inhibitory effect of podocalyxin was reversed by treatment of the cells with Arthrobacter ureafaciens sialidase, indicating that sialic acid is required for inhibition of cell adhesion. Overexpression of podocalyxin also affected transepithelial resistance and the distribution of junctional proteins in MDCK cells by an unknown mechanism that may involve interaction with the actin cytoskeleton. These results provide direct evidence that podocalyxin functions as an antiadhesin that maintains an open filtration pathway between neighboring foot processes in the glomerular epithelium by charge repulsion.

MeSH Terms
Amino Acid Sequence Animals Arthrobacter/enzymology CHO Cells Cell Adhesion/physiology Cell Aggregation/physiology Cell Line Chickens Cloning, Molecular Cricetinae Dogs Glycosylation Humans Intercellular Junctions/physiology Kidney Molecular Sequence Data Neuraminidase/metabolism Phosphorylation Rabbits Rats Receptors, Steroid/genetics,physiology Recombinant Fusion Proteins/chemistry,metabolism Sequence Alignment Sequence Homology, Amino Acid Sialoglycoproteins/chemistry,genetics,physiology Transfection
Chemicals
Receptors, Steroid Recombinant Fusion Proteins Sialoglycoproteins ecdysone receptor podocalyxin Neuraminidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Takeda T
Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA.
Go W Y
Orlando R A
Farquhar M G
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.11.9.3219
Published
2000-09-00
Pages
3219-32
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC14987
Subset
IM
Grants
NIDDK NIH HHS · R01 DK017724 · United States
NIDDK NIH HHS · R37 DK017724 · United States
NIDDK NIH HHS · DK17724 · United States
Databases
GENBANK
AF109393
External Links
PubMed source DOI: 10.1091/mbc.11.9.3219 PMC Full Text View journal details
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