Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin

K. Schwarz, M. Simons, J. Reiser, M. A. Saleem, Christian H Faul, W. Kriz, A. S. Shaw, L. B. Holzman, P. Mundel

Research output: Contribution to journalArticle

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Abstract

NPHS2 was recently identified as a gene whose mutations cause autosomal recessive steroid-resistant nephrotic syndrome. Its product, podocin, is a new member of the stomatin family, which consists of hairpin-like integral membrane proteins with intracellular NH2- and COOH-termini. Podocin is expressed in glomerular podocytes, but its subcellular distribution and interaction with other proteins are unknown. Here we show, by immunoelectron microscopy, that podocin localizes to the podocyte foot process membrane, at the insertion site of the slit diaphragm. Podocin accumulates in an oligomeric form in lipid rafts of the slit diaphragm. Moreover, GST pull-down experiments reveal that podocin associates via its COOH-terminal domain with CD2AP, a cytoplasmic binding partner of nephrin, and with nephrin itself. That podocin interacts with CD2AP and nephrin in vivo is shown by coimmunoprecipitation of these proteins from glomerular extracts. Furthermore, in vitro studies reveal direct interaction of podocin and CD2AP. Hence, as with the erythrocyte lipid raft protein stomatin, podocin is present in high-order oligomers and may serve a scaffolding function. We postulate that podocin serves in the structural organization of the slit diaphragm and the regulation of its filtration function.

Original languageEnglish
Pages (from-to)1621-1629
Number of pages9
JournalJournal of Clinical Investigation
Volume108
Issue number11
DOIs
StatePublished - Dec 29 2001
Externally publishedYes

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Diaphragm
Podocytes
Lipids
NPHS2 protein
nephrin
Proteins
Immunoelectron Microscopy
Membrane Proteins
Erythrocytes
Mutation
Membranes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Schwarz, K., Simons, M., Reiser, J., Saleem, M. A., Faul, C. H., Kriz, W., ... Mundel, P. (2001). Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. Journal of Clinical Investigation, 108(11), 1621-1629. https://doi.org/10.1172/JCI200112849

Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. / Schwarz, K.; Simons, M.; Reiser, J.; Saleem, M. A.; Faul, Christian H; Kriz, W.; Shaw, A. S.; Holzman, L. B.; Mundel, P.

In: Journal of Clinical Investigation, Vol. 108, No. 11, 29.12.2001, p. 1621-1629.

Research output: Contribution to journalArticle

Schwarz, K, Simons, M, Reiser, J, Saleem, MA, Faul, CH, Kriz, W, Shaw, AS, Holzman, LB & Mundel, P 2001, 'Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin', Journal of Clinical Investigation, vol. 108, no. 11, pp. 1621-1629. https://doi.org/10.1172/JCI200112849
Schwarz, K. ; Simons, M. ; Reiser, J. ; Saleem, M. A. ; Faul, Christian H ; Kriz, W. ; Shaw, A. S. ; Holzman, L. B. ; Mundel, P. / Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. In: Journal of Clinical Investigation. 2001 ; Vol. 108, No. 11. pp. 1621-1629.
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