Skip to main content
SpringerLink
Log in

Association of urinary macromolecules with calcium oxalate crystals induced in vitro in normal human and rat urine

  • Original Paper
  • Published:
Urological Research Aims and scope Submit manuscript

Abstract

This study was undertaken to identify proteins which are found associated with calcium oxalate crystals induced in vitro in normal human and rat urine. Crystallization was initiated by adding sodium oxalate individually to each urine sample without centrifugation and filtration. Crystals were collected and analyzed by scanning electron microscopy and X-ray diffraction. Crystal matrix proteins (CMPs) were obtained by demineralization of the crystals with ethylenediaminetetra-acetic acid (EDTA) and analyzed by western blotting technique for immunological identification. Crystals produced in human urine were found to be a mixture of calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) while those produced in rat urine were exclusively COD. CMPs extracted from crystals in human urine comprised, in addition to prothrombin-related proteins, osteopontin and albumin. However, CMPs extracted from crystals in rat urine contained only osteopontin and albumin. Prothrombin-related proteins were found only in trace amounts. In a separate experiment, rat urine samples were supplemented with COM before inducing crystallization. Similar results were observed showing that CMP contained osteopontin, albumin and trace amounts of prothrombin-related proteins. We conclude that several urinary macromolecules including not only prothrombin-related proteins, but also osteopontin and albumin, become associated with CaOx crystals. The incorporation of these proteins in growing stones is not only due to the presence of γ-carboxyglutamic acid as it was suggested for prothrombin-related proteins, but may be due to other factors such as urinary chemistry, presence of glutamic and aspartic acid residues, and calcium-binding sites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Boyce WH (1968) Organic matrix of human urinary concretions. Am J Med 45:673

    Google Scholar 

  2. Boyce WH, Garvey FK (1956) The amount and nature of the organic matrix in urinary calculi: a review. J Urol 76:213

    Google Scholar 

  3. Doyle IR, Ryall RL, Marshall VR (1991) Inclusion of proteins into calcium oxalate crystals precipitated from human urine: a highly selective phenomenon. Clin Chem 37:1589

    Google Scholar 

  4. Doyle IR, Marshall VR, Dawson CJ, Ryall RL (1995) Calcium oxalate crystal matrix extract: the most potent macromolecular inhibitor of crystal growth and aggregation yet tested in human undiluted human urine in vitro. Urol Res 23:53

    Google Scholar 

  5. Dussol B, Geider S, Lilova A, Léonetti F, Dupuy P, Daudon M, Berlan Y, Dagorn J, Verdier J (1995) Analysis of the soluble organic matrix of five morphologically different kidney stones. Evidence for a specific role of albumin in the constitution of the stone protein matrix. Urol Res 23:45

    Google Scholar 

  6. Finlayson B, Vermeulen CW, Stewart EJ (1961) Stone matrix and mucoprotein in urine. J Urol 86:355

    Google Scholar 

  7. Fraij BM (1989) Separation and identification of urinary proteins and stone-matrix proteins by mini-slab sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Clin Chem 35:658

    Google Scholar 

  8. Franzen A, Heinegard D (1985) Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochem J 232:715

    Google Scholar 

  9. Hess B (1991) The role of Tamm-Horsfall glycoprotein and nephrocalcin in calcium oxalate monohydrate crystallization processes. Scanning Micros 5:689

    Google Scholar 

  10. Hoyer JR (1994) Uropontin in urinary calcium stone formation. Miner Electrolyte Metab 20:385

    Google Scholar 

  11. Khan SR, Hackett RL (1987) Crystal-matrix relationships in experimentally induced urinary calcium oxalate monohydrate crystals, an ultrastructural study. Calcif Tissue Int 41:157

    Google Scholar 

  12. Khan SR, Hackett RL (1993) Role of organic matrix in urinary stone formation: an ultrastructural study of crystal matrix interface of calcium oxalate monohydrate stones. J Urol 150:239

    Google Scholar 

  13. Kohri K, Umekawa T, Kodama M, Katayama Y, Ishikawa Y, Takada M, Katoh Y, Kataoka K, Iguchi M, Kurita T (1990) Inhibition effect of glutamic acid and aspartic acid on calcium oxalate crystal formation. Eur Urol 17:173

    Google Scholar 

  14. Kohri K, Suzuki Y, Yoshida K, Yamamoto K, Amasaki N, Yamate T, Umekawa T, Iguchi M, Sinohara H, Kurita T (1992) Molecular cloning and sequencing of cDNA encoding urinary stone protein, which is identical to osteopontin. Biochem Biophys Res Commun 184:859

    Google Scholar 

  15. Leal J, Finlayson B (1977) Adsorption of naturally occurring polymers on calcium oxalate surfaces. Invest Urol 14:278

    Google Scholar 

  16. Lentner C (1982) Units of measurement, body fluids, composition of the body, nutrition. In: Ciba-Geity, 8th edn., Vol 1 p 57

  17. Lian JB, Prien EL, Glimcher JMJ, Gallop PM (1977) The presence of protein-bound γ-carboxyglutamic acid in calciumcontaining renal calculi. J Clin Invest 59:1151

    Google Scholar 

  18. Mckee MD, Nanci A, Khan SR (1994) Ultrastructural immunodetection of osteopontin and osteocalcin as major matrix components of urinary calculi. J Bone Miner Res 9 [Suppl] 1: S379

  19. Morrissey JH (1981) Silver stain for protein in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307

    Google Scholar 

  20. Morse RM, Resnick MI (1988) A new approach to the study of urinary macromolecules as a participant in calcium oxalate crystallization. J Urol 139:869

    Google Scholar 

  21. Morse RM, Resnick MI (1989) A study of the incorporation of urinary macromolecules onto crystals of different mineral compositions. J Urol 141:641

    Google Scholar 

  22. Shevock PN, Khan SR, Hackett R (1993) Urinary chemistry of the normal Sprague-Dawley rat. Urol Res 21:309

    Google Scholar 

  23. Shiraga H, Min W, Vandusen WJ, Clayman MD, Miner D, Terrell CH, Sherbotie JR, Foreman JW, Przysiecki C, Neilson EG, Hoyer JR (1992) Inhibition of calcium oxalate crystal growth in vitro by uropontin: another member of the aspartic acid-rich protein superfamily. Proc Natl Acad Sci USA 89:426

    Google Scholar 

  24. Stacholy J, Goldberg EP (1985) Microstructural matrix-crystal interactions in calcium oxalate monohydrate kidney stones. Scanning Electron Microsc II:781

    Google Scholar 

  25. Stapleton AMF, Ryall RM (1995) Blood coagulation proteins and urolithiasis are linked: crystal matrix protein is the F1 activation peptide of human prothrombin. Br J Urol 75:712

    Google Scholar 

  26. Stapleton AMF, Simpson RJ, Ryall RL (1993) Crystal matrix protein is related to human prothrombin. Biochem Biophys Res Comm 195:1199

    Google Scholar 

  27. Suzuki K, Moriyama M, Nakajima C, Kawamura K, Miyazawa K, Tsugawa R, Kikuchi N, Nagata K (1994) Isolation and partial characterization of crystal matrix protein as a potent inhibitor of calcium oxalate crystal aggregation: evidence of activation peptide of human prothrombin. Urol Res 22:45

    Google Scholar 

  28. Vermeulen CW, Lyon ES (1956) Mechanisms of genesis and growth of calculi. Am J Med 45:684

    Google Scholar 

  29. Warpehoski MA, Buscemi PJ, Osborn DC, Finlayson B, Goldberg EP (1981) Distribution of organic matrix in calcium oxalate calculi. Calcif Tissue Int 33:211

    Google Scholar 

  30. Worcester EM, Nakagawa Y, Wabner CL, Kumar S, Coe FL (1988) Crystal adsorption and growth slowing by nephrocalcin, albumin, and Tamm-Horsfall protein. Am J Physiol 255:1197

    Google Scholar 

  31. Worcester EM, Blumenthal SS, Beshensky AM, Lewand DL (1992) The calcium oxalate crystal growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin. J Bone Miner Res 7:1029

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cohege of Medicine, Department of Pathology and Laboratory Medicine, University of Florida, 32610-0275, Gainesville, FL, USA

    F. Atmani, F. J. Opalko & S. R. Khan

Authors
  1. F. Atmani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. J. Opalko
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. R. Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Atmani, F., Opalko, F.J. & Khan, S.R. Association of urinary macromolecules with calcium oxalate crystals induced in vitro in normal human and rat urine. Urol. Res. 24, 45–50 (1996). https://doi.org/10.1007/BF00296733

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00296733

Key words

Search

Navigation