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The calcium oxalate crystal growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin.
J Bone Miner Res. 1992 Sep; 7(9):1029-36.JB

Abstract

Urine contains proteins that inhibit the growth of calcium oxalate (CaOx) crystals and may prevent the formation of kidney stones. We have identified a potent crystal growth inhibitor in the conditioned media from primary cultures of mouse kidney cortical cells. Conditioned media, incubated with the kidney cells for 6-72 h, was assayed for crystal growth inhibition; inhibitory activity increased 15-fold by 24 h. Inhibitory activity was purified from serum-free media containing proteinase inhibitors using anion-exchange and gel-filtration chromatography. A single band of molecular weight 80,000 daltons was seen after SDS-polyacrylamide gel electrophoresis. The sequence of the N-terminal 21 amino acids of this protein matched that of osteopontin (OP), a phosphoprotein initially isolated from bone matrix. Antisera raised to fusion proteins produced by plasmids containing the N-terminal or C-terminal portions of OP cDNA also cross-reacted with the protein purified from cell culture media on western blots. The effect of the purified protein on the growth of CaOx crystals was measured using a constant composition assay. A 50% inhibition of growth occurred at a protein concentration of 0.85 micrograms/ml, and the dissociation constant of the protein with respect to CaOx crystal was 3.7 x 10(-8) M. The concentration of OP in mouse urine, measured using antibodies raised to the purified protein, was approximately 8 micrograms/ml. We conclude that OP is synthesized by kidney cortical tubule cells and functions as a crystal growth inhibitory protein in urine.

Authors+Show Affiliations

Nephrology Section, Zablocki VA Medical Center, Milwaukee.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

1414495

Citation

Worcester, E M., et al. "The Calcium Oxalate Crystal Growth Inhibitor Protein Produced By Mouse Kidney Cortical Cells in Culture Is Osteopontin." Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research, vol. 7, no. 9, 1992, pp. 1029-36.
Worcester EM, Blumenthal SS, Beshensky AM, et al. The calcium oxalate crystal growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin. J Bone Miner Res. 1992;7(9):1029-36.
Worcester, E. M., Blumenthal, S. S., Beshensky, A. M., & Lewand, D. L. (1992). The calcium oxalate crystal growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin. Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research, 7(9), 1029-36.
Worcester EM, et al. The Calcium Oxalate Crystal Growth Inhibitor Protein Produced By Mouse Kidney Cortical Cells in Culture Is Osteopontin. J Bone Miner Res. 1992;7(9):1029-36. PubMed PMID: 1414495.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The calcium oxalate crystal growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin. AU - Worcester,E M, AU - Blumenthal,S S, AU - Beshensky,A M, AU - Lewand,D L, PY - 1992/9/1/pubmed PY - 1992/9/1/medline PY - 1992/9/1/entrez SP - 1029 EP - 36 JF - Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research JO - J Bone Miner Res VL - 7 IS - 9 N2 - Urine contains proteins that inhibit the growth of calcium oxalate (CaOx) crystals and may prevent the formation of kidney stones. We have identified a potent crystal growth inhibitor in the conditioned media from primary cultures of mouse kidney cortical cells. Conditioned media, incubated with the kidney cells for 6-72 h, was assayed for crystal growth inhibition; inhibitory activity increased 15-fold by 24 h. Inhibitory activity was purified from serum-free media containing proteinase inhibitors using anion-exchange and gel-filtration chromatography. A single band of molecular weight 80,000 daltons was seen after SDS-polyacrylamide gel electrophoresis. The sequence of the N-terminal 21 amino acids of this protein matched that of osteopontin (OP), a phosphoprotein initially isolated from bone matrix. Antisera raised to fusion proteins produced by plasmids containing the N-terminal or C-terminal portions of OP cDNA also cross-reacted with the protein purified from cell culture media on western blots. The effect of the purified protein on the growth of CaOx crystals was measured using a constant composition assay. A 50% inhibition of growth occurred at a protein concentration of 0.85 micrograms/ml, and the dissociation constant of the protein with respect to CaOx crystal was 3.7 x 10(-8) M. The concentration of OP in mouse urine, measured using antibodies raised to the purified protein, was approximately 8 micrograms/ml. We conclude that OP is synthesized by kidney cortical tubule cells and functions as a crystal growth inhibitory protein in urine. SN - 0884-0431 UR - https://www.unboundmedicine.com/medline/citation/1414495/The_calcium_oxalate_crystal_growth_inhibitor_protein_produced_by_mouse_kidney_cortical_cells_in_culture_is_osteopontin_ L2 - https://doi.org/10.1002/jbmr.5650070905 DB - PRIME DP - Unbound Medicine ER -