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Bovine dentin phosphophoryn: composition and molecular weight.

W G Stetler-Stevenson, A Veis

    Biochemistry
    |August 30, 1983
    PubMed
    Summary

    Determining the molecular weight of phosphophoryn is challenging. This study isolated bovine dentin phosphophoryn (BDPP) and found its molecular weight to be significantly higher than previously reported, around 151,000-167,000.

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    Area of Science:

    • Biochemistry
    • Biomaterials Science
    • Dental Research

    Background:

    • Phosphophoryn is an acidic phosphoprotein crucial to dentin matrix.
    • Accurate molecular weight determination is hindered by protease activity and high charge density.

    Purpose of the Study:

    • To isolate and accurately determine the molecular weight of bovine dentin phosphophoryn (BDPP).
    • To overcome challenges in phosphophoryn molecular weight analysis.

    Main Methods:

    • Isolation of BDPP using a proteolysis-preventing procedure.
    • Purification via sequential precipitation, gel filtration, and anion-exchange chromatography.
    • Molecular weight determination using sedimentation equilibrium, velocity, diffusion coefficient, viscosity, and NaDodSO4 gel electrophoresis.

    Main Results:

    • Consistent molecular weight range of 151,000-167,000 for BDPP, higher than prior reports.
    • Observed anomalous behavior in non-gradient NaDodSO4 gel electrophoresis.
    • Identified a discrepancy between gel filtration (100,000 MW) and other methods, explained by non-random chain conformation in 4 M Gdn.HCl.

    Conclusions:

    • The molecular weight of BDPP is significantly higher than previously established.
    • Bovine dentin phosphophoryn does not adopt a random-chain conformation in 4 M guanidine hydrochloride.
    • This study provides a more accurate molecular weight for BDPP, crucial for understanding dentin biomineralization.

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