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Epitaxial deposition of calcium oxalate on uric acid rich stone matrix is induced by a 29 kDa protein.
Clin Chim Acta. 2006 Feb; 364(1-2):267-74.CC

Abstract

BACKGROUND

Association of macromolecules particularly the role of proteins in urolithiasis has been studied for last few centuries, but still a complete profile of stone matrix proteins that mediate co-precipitation of uric acid and calcium oxalate has not been characterized. We isolated and characterize proteins from uric acid rich stone matrix, which have oxalate binding activity.

METHODS

Matrix proteins were isolated from uric acid rich stone matrix using EDTA as a demineralizing agent. The radiolabelled solubilized proteins were fractionated with increasing ionic concentration by DEAE cellulose column chromatography to identify the oxalate binding protein. It was purified using Sephadex G-200 column chromatography. Amino acid composition was determined and monoclonal antibody was produced against the oxalate binding uric acid rich stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers urine, their oxalate binding activity assayed and cross reactivity with the produced monoclonal antibody were checked using ELISA and Western blotting.

RESULTS

Matrix on DEAE column chromatography elution yielded 3 protein peaks and they were named as fraction I, II and III among which fraction I had higher oxalate binding activity which was further purified with Sephadex G-200 column which yielded 2 protein peaks designated as Ia and Ib. Fraction Ib with molecular weight 29 kDa exhibited the maximum oxalate binding activity. Forty percent of this 29 kDa protein is comprised of basic amino acids. Monoclonal antibody (IgG1) was produced against the 29 kDa stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers, 4 protein peaks were obtained named as fraction I to IV. Among them, fraction IV having molecular weight of approximately 29 kDa cross reacted up to 85.6% with 29 kDa stone matrix protein. Moreover, urinary 29 kDa protein exhibited oxalate binding activity of 94.16 +/- 6.08 pmol/mg protein at pH 5.5.

CONCLUSION

The 29 kDa protein isolated from uric acid rich stone matrix and urine are one and the same, thereby insinuating that 29 kDa protein might play a major role in epitaxial deposition of calcium oxalate over uric acid core, consequently favoring the lithogenic events like uric acid and calcium oxalate nucleation, aggregation and retention.

Authors+Show Affiliations

Department of Medical Biochemistry, Dr. A.L. Mudaliar, Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai 600 113, India. srini@musc.eduNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16139257

Citation

Srinivasan, S, et al. "Epitaxial Deposition of Calcium Oxalate On Uric Acid Rich Stone Matrix Is Induced By a 29 kDa Protein." Clinica Chimica Acta; International Journal of Clinical Chemistry, vol. 364, no. 1-2, 2006, pp. 267-74.
Srinivasan S, Kalaiselvi P, Varalakshmi P. Epitaxial deposition of calcium oxalate on uric acid rich stone matrix is induced by a 29 kDa protein. Clin Chim Acta. 2006;364(1-2):267-74.
Srinivasan, S., Kalaiselvi, P., & Varalakshmi, P. (2006). Epitaxial deposition of calcium oxalate on uric acid rich stone matrix is induced by a 29 kDa protein. Clinica Chimica Acta; International Journal of Clinical Chemistry, 364(1-2), 267-74.
Srinivasan S, Kalaiselvi P, Varalakshmi P. Epitaxial Deposition of Calcium Oxalate On Uric Acid Rich Stone Matrix Is Induced By a 29 kDa Protein. Clin Chim Acta. 2006;364(1-2):267-74. PubMed PMID: 16139257.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Epitaxial deposition of calcium oxalate on uric acid rich stone matrix is induced by a 29 kDa protein. AU - Srinivasan,S, AU - Kalaiselvi,P, AU - Varalakshmi,P, Y1 - 2005/09/01/ PY - 2005/07/18/received PY - 2005/07/19/revised PY - 2005/07/19/accepted PY - 2005/9/6/pubmed PY - 2006/4/20/medline PY - 2005/9/6/entrez SP - 267 EP - 74 JF - Clinica chimica acta; international journal of clinical chemistry JO - Clin Chim Acta VL - 364 IS - 1-2 N2 - BACKGROUND: Association of macromolecules particularly the role of proteins in urolithiasis has been studied for last few centuries, but still a complete profile of stone matrix proteins that mediate co-precipitation of uric acid and calcium oxalate has not been characterized. We isolated and characterize proteins from uric acid rich stone matrix, which have oxalate binding activity. METHODS: Matrix proteins were isolated from uric acid rich stone matrix using EDTA as a demineralizing agent. The radiolabelled solubilized proteins were fractionated with increasing ionic concentration by DEAE cellulose column chromatography to identify the oxalate binding protein. It was purified using Sephadex G-200 column chromatography. Amino acid composition was determined and monoclonal antibody was produced against the oxalate binding uric acid rich stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers urine, their oxalate binding activity assayed and cross reactivity with the produced monoclonal antibody were checked using ELISA and Western blotting. RESULTS: Matrix on DEAE column chromatography elution yielded 3 protein peaks and they were named as fraction I, II and III among which fraction I had higher oxalate binding activity which was further purified with Sephadex G-200 column which yielded 2 protein peaks designated as Ia and Ib. Fraction Ib with molecular weight 29 kDa exhibited the maximum oxalate binding activity. Forty percent of this 29 kDa protein is comprised of basic amino acids. Monoclonal antibody (IgG1) was produced against the 29 kDa stone matrix protein. Urinary uric acid binding proteins were isolated from stone formers, 4 protein peaks were obtained named as fraction I to IV. Among them, fraction IV having molecular weight of approximately 29 kDa cross reacted up to 85.6% with 29 kDa stone matrix protein. Moreover, urinary 29 kDa protein exhibited oxalate binding activity of 94.16 +/- 6.08 pmol/mg protein at pH 5.5. CONCLUSION: The 29 kDa protein isolated from uric acid rich stone matrix and urine are one and the same, thereby insinuating that 29 kDa protein might play a major role in epitaxial deposition of calcium oxalate over uric acid core, consequently favoring the lithogenic events like uric acid and calcium oxalate nucleation, aggregation and retention. SN - 0009-8981 UR - https://www.unboundmedicine.com/medline/citation/16139257/Epitaxial_deposition_of_calcium_oxalate_on_uric_acid_rich_stone_matrix_is_induced_by_a_29_kDa_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0009-8981(05)00462-6 DB - PRIME DP - Unbound Medicine ER -