Proteomic analysis of altered proteins in distal renal tubular cells in response to calcium oxalate monohydrate crystal adhesion: Implications for kidney stone disease

2008 ◽  
Vol 2 (7-8) ◽  
pp. 1099-1109 ◽  
Author(s):  
Theptida Semangoen ◽  
Supachok Sinchaikul ◽  
Shui-Tein Chen ◽  
Visith Thongboonkerd
Keyword(s):  
Proteomic Analysis ◽  
Kidney Stone ◽  
Calcium Oxalate Monohydrate ◽  
Stone Disease ◽  
Kidney Stone Disease ◽  
Tubular Cells ◽  
Renal Tubular Cells ◽  
Crystal Adhesion ◽  
Renal Tubular ◽  
Altered Proteins

P0131HIGH-DOSE URIC ACID ALTERS CELLULAR PROTEOME, INCREASES INTRACELLULAR ATP, ENHANCES TISSUE REPAIR CAPABILITY AND INCREASES CALCIUM OXALATE CRYSTAL-BINDING CAPABILITY OF RENAL TUBULAR CELLS: IMPLICATIONS TO HYPERURICOSURIA-INDUCED KIDNEY STONE DISEASE

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Suchitra Sutthimethakorn ◽  
Visith Thongboonkerd
Keyword(s):  
Uric Acid ◽  
Western Blotting ◽  
Kidney Stone ◽  
Quantitative Proteomics ◽  
Stone Disease ◽  
Kidney Stone Disease ◽  
Tubular Cells ◽  
Renal Tubular Cells ◽  
Binding Capability ◽  
Renal Tubular

Abstract Background and Aims Hyperuricosuria is well documented to get involved in the pathogenesis of kidney stone disease, especially uric acid (UA) and calcium oxalate (CaOx) types. Nevertheless, hyperuricosuria-induced alterations in renal tubular cells and cascade mechanisms that subsequently trigger kidney stone formation remained largely unknown. This study aimed to examine changes in cellular proteome and function in renal tubular cells after treatment with high-dose UA. Method MDCK cells were incubated with 3.5 mM uric acid for 48-h. Cellular proteins were extracted and subjected to quantitative proteomics analysis using 2-D PAGE followed by nanoLC-ESI-ETD MS/MS. The proteome data were confirmed by Western blotting, whereas functional analyses were performed using various assays. Results Quantitative proteomics analysis revealed significant changes in levels of 22 proteins in the UA-treated cells that were related to ATPase activity, mitochondrial transport, cell adhesion and molecule binding, cell projection organization and maintenance of location network, metabolic process, response to unfold proteins, and vesicle-mediated transport. Western blotting confirmed changes in levels of cellular proteins identified by quantitative proteomics. Functional assays revealed an increase in intracellular ATP level and enhancement of tissue repair capability in the UA-treated cells. Interestingly, isolation/purification of apical membranes followed by Western blotting showed the increased levels of HSP70 and HSP90 (the known receptors for CaOx crystals) on apical membranes of the UA-treated cells. CaOx crystal-cell adhesion assay revealed significant increase in CaOx-binding capability of the UA-treated cells, whereas neutralization of the surface HSP70 and HSP90 using their specific monoclonal antibodies caused significant reduction in such binding capability (even below the basal level). Conclusion These findings highlighted changes in renal tubular cells in response to high-dose UA that might be related to the pathogenic mechanisms of kidney stone disease associated with hyperuricosuria.

scholarly journals Proteomic Analysis of Calcium Oxalate Monohydrate Crystal-Induced Cytotoxicity in Distal Renal Tubular Cells

2008 ◽  
Vol 7 (11) ◽  
pp. 4689-4700 ◽  
Author(s):  
Visith Thongboonkerd ◽  
Theptida Semangoen ◽  
Supachok Sinchaikul ◽  
Shui-Tein Chen
Keyword(s):  
Calcium Oxalate ◽  
Proteomic Analysis ◽  
Calcium Oxalate Monohydrate ◽  
Tubular Cells ◽  
Renal Tubular Cells ◽  
Renal Tubular
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