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PMID: 11457882 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Loss of glomerular foot processes is associated with uncoupling of podocalyxin from the actin cytoskeleton.

The Journal of clinical investigation ·Vol. 108 ·No. 2 ·2001-07-00 ·Pages 289-301

Takeda T, McQuistan T, Orlando RA, Farquhar MG

Abstract

Podocalyxin (PC), the major sialoprotein of glomerular epithelial cells (GECs), helps maintain the characteristic architecture of the foot processes and the patency of the filtration slits. PC associates with actin via ezrin, a member of the ERM family of cytoskeletal linker proteins. Here we show that PC is linked to ezrin and the actin cytoskeleton via Na(+)/H(+)-exchanger regulatory factor 2 (NHERF2), a scaffold protein containing two PDZ (PSD-95/Dlg/ZO-1) domains and an ERM-binding region. The cytoplasmic tail of PC contains a C-terminal PDZ-binding motif (DTHL) that binds to the second PDZ domain of NHERF2 in yeast two-hybrid and in vitro pull-down assays. By immunocytochemistry NHERF2 colocalizes with PC and ezrin along the apical domain of the GEC plasma membrane. NHERF2 and ezrin form a multimeric complex with PC, as they coimmunoprecipitate with PC. The PC/NHERF2/ezrin complex interacts with the actin cytoskeleton, and this interaction is disrupted in GECs from puromycin aminonucleoside-, protamine sulfate-, or sialidase-treated rats, which show a dramatic loss of foot processes, comparable to that seen in the nephrotic syndrome. Thus NHERF2 appears to function as a scaffold protein linking PC to ezrin and the actin cytoskeleton. PC/NHERF2/ezrin/actin interactions are disrupted in pathologic conditions associated with changes in GEC foot processes, indicating their importance for maintaining the unique organization of this epithelium.

MeSH Terms
Actins/analysis,chemistry,metabolism Animals Binding Sites Cell Membrane/metabolism Cells, Cultured Cytoskeletal Proteins Cytoskeleton/chemistry,metabolism DNA, Complementary/analysis Epithelial Cells/metabolism Immunohistochemistry Kidney Glomerulus/metabolism,pathology,ultrastructure Male Molecular Sequence Data Nephrotic Syndrome/metabolism,pathology Phosphoproteins/analysis,chemistry,genetics Rats Rats, Sprague-Dawley Sialoglycoproteins/analysis,chemistry,metabolism Sodium-Hydrogen Exchangers/chemistry,metabolism
Chemicals
Actins Cytoskeletal Proteins DNA, Complementary Phosphoproteins Sialoglycoproteins Sodium-Hydrogen Exchangers ezrin podocalyxin sodium-hydrogen exchanger regulatory factor
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, USA.
McQuistan T
Orlando R A
Farquhar M G
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI12539
Published
2001-07-00
Pages
289-301
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC203027
Subset
IM
Grants
NIDDK NIH HHS · R01 DK017724 · United States
NIDDK NIH HHS · R37 DK017724 · United States
NIDDK NIH HHS · DK-17724 · United States
Databases
GENBANK
AF294257
Corrections
CommentIn
PMID: 11457870
External Links
PubMed source DOI: 10.1172/JCI12539 PMC Full Text View journal details
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