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

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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.

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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

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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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Authors and Affiliations

  1. Department of Chemistry, University of Cape Town, Private Bag, Rondebosch, Cape Town, 7701, South Africa

    Dawn Webber & Allen L. Rodgers

  2. Division of Medical Biochemistry and Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Observatory, Cape Town, 7925, South Africa

    Edward D. Sturrock

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  1. Dawn Webber
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  2. Allen L. Rodgers
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  3. Edward D. Sturrock
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Correspondence to Dawn Webber.

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Webber, D., Rodgers, A.L. & Sturrock, E.D. Glycosylation of prothrombin fragment 1 governs calcium oxalate crystal nucleation and aggregation, but not crystal growth. Urol Res 35, 277–285 (2007). https://doi.org/10.1007/s00240-007-0119-z

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  • DOI: https://doi.org/10.1007/s00240-007-0119-z

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