Bovine Dentin Phosphophoryn: Composition and Molecular Weight

W. G. Stetler-Stevenson, Arthur Veis*

*Corresponding author for this work

Research output: Contribution to journalArticle

94 Scopus citations

Abstract

The molecular weight of phosphophoryn, an acidic phosphoprotein unique to dentin matrix, has been difficult to determine because of a combination of neutral protease activities in this tissue and the intrinsic high charge density of the molecule. In this study, bovine dentin phosphophoryn (BDPP) was isolated by a procedure designed to prevent proteolysis. Bovine unerupted third molar powder was demineralized by ethylenediaminetetraacetic acid (EDTA). The EDTA-soluble phosphophoryn fraction was isolated and purified by sequential calcium chloride precipitation, gel filtration in sodium dodecyl sulfate (NaDodSO4) containing buffer, anion-exchange chromatography, and finally gel filtration in 4 M guanidine hydrochloride (4 M Gdn·HCl) buffer. Sedimentation equilibrium, sedimentation velocity, and diffusion coefficient data, viscosity studies in a high ionic strength buffer, and NaDodSO4 gradient gel electrophoresis data gave consistent results for the molecular weight of BDPP, all being in the range of 151 000–167 000. This range is much higher than any previously reported value. An anomalous behavior was observed in nongradient NaDodSO4 gel electrophoresis. Dissociative analytical gel filtration chromatography in 4 M Gdn·HCl gave a molecular weight value of 100 000. This discrepancy was resolved by studying the viscosity of BDPP in 4 M Gdn·HCl which showed BDPP does not assume a true random-chain conformation in this solvent.

Original languageEnglish (US)
Pages (from-to)4326-4335
Number of pages10
JournalBiochemistry
Volume22
Issue number18
DOIs
StatePublished - Jan 1 1983

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Bovine Dentin Phosphophoryn: Composition and Molecular Weight'. Together they form a unique fingerprint.

  • Cite this