Annexin II is present on renal epithelial cells and binds calcium oxalate monohydrate crystals

Vivek Kumar, Gerard Farell, Sergio Deganello, John C Lieske

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Abstract

Attachment of newly formed crystals to renal epithelial cells appears to be a critical step in the development of kidney stones. The current study was undertaken to identify potential calcium oxalate monohydrate (COM) crystal-binding proteins on the surface of renal tubular cells. Apical membranes were prepared from confluent monolayers of renal epithelial cells (MDCKI line), and COM crystal affinity was used to isolate crystal-binding proteins that were then subjected to electrophoresis and electroblotting. Microsequencing of the most prominent COM crystal-binding protein (M r of 37 kD) identified it as annexin II (Ax-II). When exposed proteins on the surface of intact'monolayers were biotinylated and then isolated using streptavidin agarose beads, Ax-II was detected, suggesting that at least a portion is exposed on the apical cell surface. Ax-II was not completely extracted by 0.1 M N 2CO 3, suggesting that at least a portion of cellular Ax-II is an intrinsic membrane-bound protein. Using confocal immunofluorescence microscopy, Ax-II was visualized together with Caveolin-1 (Cav-1) on the apical membrane of intact MDCKI cells. Cells pretreated with a monoclonal anti-Ax-II antibody bound significantly fewer COM crystals, whereas anti-LDL receptor antibody did not decrease COM binding, further suggesting a functional role for Ax-II during adhesion of crystals to intact cells. These results suggest that Ax-II avidly binds COM crystals and is present on the apical surface of MDCKI cells. Therefore, in the intact nephron, Ax-II could mediate adhesion of COM crystals to cells, and altered exposure of Ax-II on the surface of renal tubular cells could promote crystal retention and possibly kidney stone formation.

Original languageEnglish (US)
Pages (from-to)289-297
Number of pages9
JournalJournal of the American Society of Nephrology
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2003

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Annexin A2
Calcium Oxalate
Epithelial Cells
Kidney
Carrier Proteins
Kidney Calculi
Membrane Proteins
Caveolin 1
Membranes
Antibodies
LDL Receptors
Nephrons
Fluorescence Microscopy
Confocal Microscopy
Electrophoresis

ASJC Scopus subject areas

  • Nephrology

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Annexin II is present on renal epithelial cells and binds calcium oxalate monohydrate crystals. / Kumar, Vivek; Farell, Gerard; Deganello, Sergio; Lieske, John C.

In: Journal of the American Society of Nephrology, Vol. 14, No. 2, 01.02.2003, p. 289-297.

Research output: Contribution to journalArticle

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