Urological Research

, Volume 35, Issue 6, pp 277–285 | Cite as

Glycosylation of prothrombin fragment 1 governs calcium oxalate crystal nucleation and aggregation, but not crystal growth

  • Dawn Webber
  • Allen L. Rodgers
  • Edward D. Sturrock
Original Paper

Abstract

Urinary glycoproteins play an important role in the modulation of calcium oxalate crystallisation. In several cases, this has been attributed to glycosylation of the proteins as evidenced by urinary prothrombin fragment 1 where there is a correlation between sialylation and calcium oxalate kidney stone disease. In the present study, plasma-derived prothrombin fragment 1 (PTF1) was enzymatically modified in order to generate its asialo and aglyco forms. The parent glycoprotein and its two glycoforms were used in calcium oxalate crystallisation studies to assess the role of the carbohydrate moeity in PTF1’s potent inhibitory activity. The glycans inhibited crystal aggregation and promoted crystal nucleation, but had no effect on crystal growth. The terminal sialic acid residues had a small effect on inhibition of crystal aggregation whereas they contributed significantly to promotion of nucleation. These results indicate that glycosylation of PTF1 governs calcium oxalate crystal nucleation and aggregation but it does not affect the protein’s role in inhibiting crystal growth. Since promotion of nucleation and inhibition of aggregation are both regarded as protective mechanisms against calcium oxalate urinary stone formation, the kringle domain on which the glycans are located is implicated in PTF1’s inhibitory activity. It is speculated that modifications in the glycosylation of urinary PTF1 in stone-formers may regulate its capacity to protect against calcium urolithiasis.

Keywords

Calcium oxalate Crystallization Glycosylation Kidney stones Protein Prothrombin fragment 1 

Abbreviations

agPTF1

Aglyco prothrombin fragment 1

asPTF1

Asialo prothrombin fragment 1

CaOx

Calcium oxalate

Gla

γ-carboxyglutamic acid

gPTF1

Glyco prothrombin fragment 1

IN

Inhibition of nucleation

IG

Inhibition of growth

MALDI-TOF

Matrix-assisted laser-desorption-ionisation time-of-flight

PT

Prothrombin

PTF1

Prothrombin fragment 1

SEM

Standard error of mean

SU

Synthetic urine

UPTF1

Urinary prothrombin fragment 1

Notes

Acknowledgments

The authors wish to thank the South African Medical Research Council, the South African National Research Foundation, the South African Urological Association, the University of Cape Town and the Volkswagen Stiftung for financial support. Thanks are also accorded to Mr Ian Durbach for assistance with the statistical analysis.

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Dawn Webber
    • 1
  • Allen L. Rodgers
    • 1
  • Edward D. Sturrock
    • 2
  1. 1.Department of ChemistryUniversity of Cape TownCape TownSouth Africa
  2. 2.Division of Medical Biochemistry and Institute of Infectious Disease and Molecular MedicineUniversity of Cape TownCape TownSouth Africa

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