<Review>Urinary stone : matrix protein ; osteopontin and calprotectin

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The chemical nature of the urinary stone protein is poorly understood. A cDNA of urinary calcium oxalate stone (CaOx) protein was extracted with EDTA. cDNA sequences showed complete identity between urinary stoe protein and human osteopontin. Ostepontin protein was detected by staining with Stains-All, which specifically stains phosphoproteins, and by digestion with the highly specific protease thrombin, demonstrating that urinary CaOx consists of osteopontin protein. In situ hybridization was used to detect osteopontin mRNA in the kidney. In control normal rats, distal tubular cells were sporadically positive, and proximal tubular cells and glomeruli were negative for osteopontin mRNA. A rat model of stone formation was induced with glyoxylic acid. In stone-forming rats, staining of distal tubular cells was remarkably increased, but proximal tubular cells and glomeruli were still negative. Immunostaining for the osteopontin protein also revealed that epithelial cells of distal tubules were weakly positive in control rats and significantly increased in stone-forming rats, although proximal tubular cells and glomeruli were negative. Northern blot analysis showed a significant increase of osteopontin mRNA in stone-forming rats in proportion to the dosage and the duration of the stone-inducing drugs. These results show that osteopontin in the kidney is presumably involved in urinary stone formation as the important stone matrix. Calprotectin an another important matrix protein in the urinary stone was also described in this review. Soluble organic matrix was extracted with EDTA from calcium oxalate monohydrate urinary stone, purified by SDS-polyacryamide gel electrophoresis and transferred to polyvinylidene difluoride membranes for NH_2-terminal amino acid sequence analysis. A protein at 30 kDa showed complete homology with calprotectin (20 amino acids in the NH_2-terminal), so we termed this a Calprotectin-like protein. Calprotectin which was extracted from human granulocytes had inhibitory activity in calcium CaOx crystal growth in virto. These results suggested that calprotectin-like protein detected in CaOx is a potent inhibitor of crystal growth and may therefore be importaint in the etiology of CaOx formation.
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<Review>Urinary stone : matrix protein ; osteopontin and calprotectin

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