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Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin

We study the effect of isoforms of osteopontin (OPN) on the nucleation and growth of crystals from a supersaturated solution of calcium and phosphate ions. Dynamic light scattering is used to monitor the size of the precipitating particles and to provide information about their concentration. At the...

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Autores principales: de Bruyn, John R., Goiko, Maria, Mozaffari, Maryam, Bator, Daniel, Dauphinee, Ron L., Liao, Yinyin, Flemming, Roberta L., Bramble, Michael S., Hunter, Graeme K., Goldberg, Harvey A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3572982/
https://www.ncbi.nlm.nih.gov/pubmed/23457612
http://dx.doi.org/10.1371/journal.pone.0056764
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author de Bruyn, John R.
Goiko, Maria
Mozaffari, Maryam
Bator, Daniel
Dauphinee, Ron L.
Liao, Yinyin
Flemming, Roberta L.
Bramble, Michael S.
Hunter, Graeme K.
Goldberg, Harvey A.
author_facet de Bruyn, John R.
Goiko, Maria
Mozaffari, Maryam
Bator, Daniel
Dauphinee, Ron L.
Liao, Yinyin
Flemming, Roberta L.
Bramble, Michael S.
Hunter, Graeme K.
Goldberg, Harvey A.
author_sort de Bruyn, John R.
collection PubMed
description We study the effect of isoforms of osteopontin (OPN) on the nucleation and growth of crystals from a supersaturated solution of calcium and phosphate ions. Dynamic light scattering is used to monitor the size of the precipitating particles and to provide information about their concentration. At the ion concentrations studied, immediate precipitation was observed in control experiments with no osteopontin in the solution, and the size of the precipitating particles increased steadily with time. The precipitate was identified as hydroxyapatite by X-ray diffraction. Addition of native osteopontin (nOPN) extracted from rat bone caused a delay in the onset of precipitation and reduced the number of particles that formed, but the few particles that did form grew to a larger size than in the absence of the protein. Recombinant osteopontin (rOPN), which lacks phosphorylation, caused no delay in initial calcium phosphate precipitation but severely slowed crystal growth, suggesting that rOPN inhibits growth but not nucleation. rOPN treated with protein kinase CK2 to phosphorylate the molecule (p-rOPN) produced an effect similar to that of nOPN, but at higher protein concentrations and to a lesser extent. These results suggest that phosphorylations are critical to OPN’s ability to inhibit nucleation, whereas the growth of the hydroxyapatite crystals is effectively controlled by the highly acidic OPN polypeptide. This work also demonstrates that dynamic light scattering can be a powerful tool for delineating the mechanism of protein modulation of mineral formation.
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spelling pubmed-35729822013-03-01 Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin de Bruyn, John R. Goiko, Maria Mozaffari, Maryam Bator, Daniel Dauphinee, Ron L. Liao, Yinyin Flemming, Roberta L. Bramble, Michael S. Hunter, Graeme K. Goldberg, Harvey A. PLoS One Research Article We study the effect of isoforms of osteopontin (OPN) on the nucleation and growth of crystals from a supersaturated solution of calcium and phosphate ions. Dynamic light scattering is used to monitor the size of the precipitating particles and to provide information about their concentration. At the ion concentrations studied, immediate precipitation was observed in control experiments with no osteopontin in the solution, and the size of the precipitating particles increased steadily with time. The precipitate was identified as hydroxyapatite by X-ray diffraction. Addition of native osteopontin (nOPN) extracted from rat bone caused a delay in the onset of precipitation and reduced the number of particles that formed, but the few particles that did form grew to a larger size than in the absence of the protein. Recombinant osteopontin (rOPN), which lacks phosphorylation, caused no delay in initial calcium phosphate precipitation but severely slowed crystal growth, suggesting that rOPN inhibits growth but not nucleation. rOPN treated with protein kinase CK2 to phosphorylate the molecule (p-rOPN) produced an effect similar to that of nOPN, but at higher protein concentrations and to a lesser extent. These results suggest that phosphorylations are critical to OPN’s ability to inhibit nucleation, whereas the growth of the hydroxyapatite crystals is effectively controlled by the highly acidic OPN polypeptide. This work also demonstrates that dynamic light scattering can be a powerful tool for delineating the mechanism of protein modulation of mineral formation. Public Library of Science 2013-02-14 /pmc/articles/PMC3572982/ /pubmed/23457612 http://dx.doi.org/10.1371/journal.pone.0056764 Text en © 2013 de Bruyn et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
de Bruyn, John R.
Goiko, Maria
Mozaffari, Maryam
Bator, Daniel
Dauphinee, Ron L.
Liao, Yinyin
Flemming, Roberta L.
Bramble, Michael S.
Hunter, Graeme K.
Goldberg, Harvey A.
Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title_full Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title_fullStr Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title_full_unstemmed Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title_short Dynamic Light Scattering Study of Inhibition of Nucleation and Growth of Hydroxyapatite Crystals by Osteopontin
title_sort dynamic light scattering study of inhibition of nucleation and growth of hydroxyapatite crystals by osteopontin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3572982/
https://www.ncbi.nlm.nih.gov/pubmed/23457612
http://dx.doi.org/10.1371/journal.pone.0056764
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