Molecular Characteristics of Uronic-Acid-Rich Protein, a Strong Inhibitor of Calcium Oxalate Crystallization In Vitro

Atmani, Fouad; Lacour, Bernard; Strecker, G; Parvy, P; Drueke, Tilman B.; Daudon, Michel

doi:10.1006/bbrc.1993.1338

Cited by 38 publications

(27 citation statements)

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“…2,13 There are many reports about the proteins extracted from human urine and the matrices of urinary stones which exhibit inhibitory activity on calcium oxalate crystal nucleation, growth, and aggregation. 4,5,14,15 Nephrocalcin is a known glycoprotein which exhibits a strong inhibitory effect on nucleation, growth, and aggregation of COM crystals. 16 Recently, it has been reported that these substances also have some effect on crystal-cell interaction.…”

Section: Discussionmentioning

confidence: 99%

“…Calcium oxalate, calcium phosphate and uric acid crystal preparation 14 C-Radiolabeled COM crystals were prepared as described previously. 10 Briefly, 10 mmol CaCl 2 was added to an equal volume of 10 mmol sodium oxalate (2 µ Ci, 14 C-oxalic acid) in artificial urine buffer, pH 6.0.…”

Section: Electrophoretic Analysismentioning

confidence: 99%

“…10 Briefly, 10 mmol CaCl 2 was added to an equal volume of 10 mmol sodium oxalate (2 µ Ci, 14 C-oxalic acid) in artificial urine buffer, pH 6.0.…”

Section: Electrophoretic Analysismentioning

confidence: 99%

“…The coverslips and MDCK cells were washed twice with artificial urine (pH 6.0) and then incubated with a well-dispersed 0.5 mg/mL 14 C-COM, 14 C-COD, 45 Cabrushite, 45 Ca-apatite, and 14 C-uric acid crystal suspension for 20 min at 37 ° C, respectively. Non-binding crystals were washed free by adhering the coverslips to a microscope slide with a dot of white petroleum jelly and dipping the slide for one minute into a slowly stirred beaker of artificial urine at room temperature.…”

Section: Electrophoretic Analysismentioning

confidence: 99%

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Calcium oxalate monohydrate crystal binding substance produced from Madin‐Darby canine kidney cells

et al. 2002

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Background : The interaction between kidney urothelium and crystals is a critical event in the growth of renal calculi. When studying calcium oxalate monohydrate (COM) crystal binding to MadinDarby canine kidney (MDCK) cells in culture, we observed that crystals also attached to areas on the coverslips devoid of cells. This phenomenon could be the result of substances produced by the cells that adhere to the glass and subsequently bind COM crystals. We investigated the characteristics of this COM binding substance. Methods : Media was collected from cultures of MDCK cells (conditioned media) and proteins were separated by high performance liquid chromatography. The molecular weights and purity of isolated proteins were determined by polyacrylamide gel electrophoresis. The conditioned media and each separated fraction were applied to glass and to MDCK cells and COM-binding ability determined using 14 C-labeled crystals. The binding of radio-labelled calcium oxalate dihydrate, brushite, uric acid, and apatite to coverslips were also studied. Results : Fourteen times more COM bound to coverslips incubated with conditioned media than those with control media. The molecular weight of the protein bound to the glass was determined to be 200 kDa. The COM crystals binding to this protein was 1.5 µ g/ng. Other crystals bound to a lesser extent. The incubation of cells with this protein inhibited COM binding by 39%. Conclusion : The MDCK cells produce a 200-kDa protein that has a high binding affinity for COM crystals. This protein binds to glass and is responsible for crystal binding to areas devoid of cells. This protein also has an inhibitory effect on COM binding to MDCK cells in culture.

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Section: Discussionmentioning

confidence: 99%

Section: Electrophoretic Analysismentioning

confidence: 99%

“…10 Briefly, 10 mmol CaCl 2 was added to an equal volume of 10 mmol sodium oxalate (2 µ Ci, 14 C-oxalic acid) in artificial urine buffer, pH 6.0.…”

Section: Electrophoretic Analysismentioning

confidence: 99%

Section: Electrophoretic Analysismentioning

confidence: 99%

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Calcium oxalate monohydrate crystal binding substance produced from Madin‐Darby canine kidney cells

et al. 2002

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“…They concluded that a predominant proteoglycan they had purified was urinary trypsin inhibitor, one of the interalpha-trypsin inhibitor (ITI) proteoglycan family. Atmani et al 25 have isolated a glycoprotein with uronic acid chains. It had a molecular weight of 35 kDa and inhibited CaOx crystal growth in an in vitro crystallization system.…”

Section: Discussionmentioning

confidence: 99%

Proteoglycan core protein in human urine and its possible role on calcium oxalate urolithiasis

Yoshimura

Miyake²,

Tsujihata³

et al. 1999

Int J of Urology

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Background: There are only a few papers reporting on the role of proteoglycan core protein in calcium oxalate stone formation. The present study was carried out to investigate the role of core protein of proteoglycan in human urine on calcium oxalate (CaOx) crystallization. Methods: Proteoglycans were collected from whole human urine. The covalently bound glycosaminoglycans (GAG) of proteoglycans were then digested by GAG lyase. The inhibitory activity on CaOx crystal growth in vitro was measured before and after enzyme digestion of proteoglycans. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the core protein of proteoglycans and the analysis of amino acid sequence were performed. Results:The core protein showed significant inhibitory activity on CaOx crystal growth, which scarcely changed when compared with that of proteoglycans before enzyme digestion. The SDS-PAGE revealed that the core protein was a single unit with a molecular weight of 26 kDa and amino acid sequencing demonstrated high homology to interalpha-trypsin inhibitor (ITI) light chain (bikunin) with Kunitz inhibitor domain as a core protein. Conclusions:The results suggested that human urine contains proteoglycans and a major part of them is ITI light chain (bikunin). The Kunitz inhibitor domain, a core protein of bikunin, has significant inhibitory activity on CaOx crystallization without GAG bound covalently to the core protein.

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Modulators of Crystallization of Stone Salts

Khan¹,

Kok²

Urinary Stone Disease

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Molecular Characteristics of Uronic-Acid-Rich Protein, a Strong Inhibitor of Calcium Oxalate Crystallization In Vitro

Cited by 38 publications

References 0 publications

Calcium oxalate monohydrate crystal binding substance produced from Madin‐Darby canine kidney cells

Calcium oxalate monohydrate crystal binding substance produced from Madin‐Darby canine kidney cells

Proteoglycan core protein in human urine and its possible role on calcium oxalate urolithiasis

Modulators of Crystallization of Stone Salts

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