Purification, composition, and31P NMR spectroscopic properties of a noncollagenous phosphoprotein isolated from chicken bone matrix

Lee, Sandra L.; Glimcher, Melvin J.

doi:10.1007/bf02409461

Cited by 64 publications

(22 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These studies were rapidly followed by the isolation of EDTA-and HC1-soluble phosphoproteins from dentin (Veis and Perry, 1967;Butler et al, 1972Butler et al, , 1976 and bone Glimcher, 1972, 1973;Lee and Glimcher, 1981;Cohen-Solal et al, 1978Uchiyama et al, 1986) and in a Z Z the other normally and pathologically calcified vertebrae tissues studied (Glimcher, 1979;Glimcher et al, 1979aGlimcher et al, ,c, 1981Lian et al, 1977Lian et al, , 1983.…”

Section: Sites Of Nucleationmentioning

confidence: 99%

Mechanism of calcification: Role of collagen fibrils and collagen‐phosphoprotein complexes in vitro and in vivo

Glimcher¹

1989

Anat. Rec.

Self Cite

307

155

View full text Add to dashboard Cite

Samples of decalcified chicken bone together with varying concentrations of phosphoproteins from bone or egg yolk (phosvitin) were used in vitro as heterogenous nucleators for the induction of Ca-P apatite crystals. The lag time between exposure of the collagen-phosphoprotein complexes and the time nucleation of crystals occurred decreased as the concentration of Ser(P) and Thr(P) increased. Enzymatic cleavage of the phosphate groups by wheat germ and phosphatase reversed this effort, indicating that the phosphate group per se principally facilitated the nucleation of Ca-P crystals by the phosphoprotein complex and collagen.

show abstract

Section: Sites Of Nucleationmentioning

confidence: 99%

Mechanism of calcification: Role of collagen fibrils and collagen‐phosphoprotein complexes in vitro and in vivo

Glimcher¹

1989

Anat. Rec.

Self Cite

307

155

View full text Add to dashboard Cite

show abstract

“…Their ubiquitous nature and other considerations have led to the hypothesis that this class of protein may play a crucial role in initiating or controlling the spatial distribution of mineral in the extracellular matrix of dentin and bone (Veis, 1978;Veis and Sabsay, 1983;Glimcher, 1976Glimcher, , 1989. For chicken bone, numerous phosphorylated polypeptides, ranging in molecular weight from 6 kDa to 150 kDa and containing both 0-phosphoserine (Ser-P) and 0phosphothreonine (Thr-P) residues, have been extracted and purified to homogeneity (Spector and Glimcher, 1972;Cohen-Solal et al, 1978;Lee and Glimcher, 1981;Uchiyama et al, 1986). The chicken bone phosphoproteins have been shown to be synthesized and secreted by osteoblasts in bone tissue in vivo and by cultured chicken bone organ cultures and cultured osteoblasts in vitro Gotoh et al, 1983Landis et al, 1984;Gerstenfeld et al, 1989).…”

mentioning

confidence: 99%

Expression and ultrastructural immunolocalization of a major 66 kDa phosphoprotein synthesized by chicken osteoblasts during mineralization in vitro

et al. 1990

View full text Add to dashboard Cite

Embryonic chicken osteoblasts cultured over a 30 day period were used as a model system for studying the expression of bone phosphoproteins during cellular differentiation and the possible role of these proteins in extracellular matrix mineralization. Accumulation of total phosphoprotein in the cultures, as determined by O-phosphoserine (Ser-P) and O-phosphothreonine (Thr-P) amino acid analysis, revealed a greater than 10-fold increase over the 30 day period. Total phosphoprotein synthesis, as assessed by (32P)-, (3H)-Ser-P, and (14C)-Thr-P protein labeling, showed the highest levels concurrent with initial mineral deposition within the matrix. The major phosphoprotein present in chicken bones and synthesized by the cultured osteoblasts had a molecular weight of approximately 66 kDa. This 66 kDa bone phosphoprotein (66 kDa BPP) was purified to homogeneity and was used for antibody production. Application of this antibody in Western blot analysis revealed that 66 kDa BPP was present only in protein extracts of mineralizing cultured osteoblasts and was absent in cultures of non-mineralizing chondrocytes, myoblasts, and tendon fibroblasts. The 66 kDa BPP in vitro accumulated continuously in the extracellular matrix in a manner that paralleled both phosphoprotein synthesis and total phospho-amino acid production. A comparison of the results obtained in vitro to those from developing embryonic tibiae in vivo demonstrated a similar qualitative and temporal expression of phosphoprotein and a continual accumulation of 66 kDa BPP in the matrix with advancing mineralization and developmental age. Ultrastructural immunocytochemistry using the 66 kDa BPP antibody and the protein A-gold technique revealed specific immunolabeling over electron-dense regions of mineralization in the cultures that appeared identical to the distribution of labeling observed in vivo (McKee et al.: Connect. Tissue Res., 21:21-29, 1989; Anat. Rec., 228:77-92, 1990). These results demonstrate that this major 66 kDa BPP was expressed concurrently with other differentiated osteoblast functions and suggests that it may play a role in the initiation or regulation of mineralization.

show abstract

“…The phosphoproteins of 0 1990 WILEY-LISS. INC chicken bone have been extensively examined (reviewed by Glimcher, 1984;Uchiyama et al, 1986), and numerous phosphorylated polypeptides, ranging in molecular weight from 6 kDa to 150 kDa and containing both Ser-P and Thr-P residues, have been isolated (Spector and Glimcher, 1972;Cohen-Solal et al, 1978;Lee and Glimcher, 1981;Uchiyama et al, 1986). Recently, a major -66 kDa bone phosphoprotein (66 kDa BPP), present in post-natal chicken bone (Uchiyama et al, 1986) and synthesized and secreted by chicken osteoblasts in cell culture (Gerstenfeld et al, 1989, has been isolated and purified (Gerstenfeld et al, 1989Gotoh et al, 1990).…”

mentioning

confidence: 99%

Developmental appearance and ultrastructural immunolocalization of a major 66 kDa phosphoprotein in embryonic and post‐natal chicken bone

et al. 1990

View full text Add to dashboard Cite

Biochemical analyses and immunocytochemistry were used to examine the developmental appearance of a major approximately 66 kDa bone phosphoprotein (66 kDa BPP) in the mid-diaphyseal region of embryonic and post-natal chicken tibiae in vivo. Total protein and O-phosphoserine (Ser-P) and O-phosphothreonine (Thr-P) content of 8-, 12-, and 18-day embryonic, and 4-wk post-natal chicken tibiae were determined by amino acid analysis. Similar bone samples were carried through a wide variety of tissue-processing regimes including different protocols for fixation, decalcification, dehydration, and embedding prior to electron microscopy. For immunocytochemistry, tissue sections were incubated with a polyclonal antibody raised in rabbits against 66 kDa BPP, and the antigen was revealed by the high-resolution protein A-gold technique. Amino acid analysis, Western blotting, and immunocytochemistry all showed the presence and increasing concentration of bone phosphoprotein with advancing developmental age. Immunogold labeling was observed over osteoblasts and mineral deposits throughout the bone with the most intense reaction occurring at the mineralization front in embryonic tibiae. Electron probe X-ray microanalysis confirmed the association of 66 kDa BPP with mineral. The levels of phosphoprotein in the tissue were directly correlated with increasing degrees of mineralization. These observations are consistent with previous proposals suggesting that phosphoproteins may play a significant role in the calcification of bone matrix.

show abstract

Purification, composition, and31P NMR spectroscopic properties of a noncollagenous phosphoprotein isolated from chicken bone matrix

Cited by 64 publications

References 30 publications

Mechanism of calcification: Role of collagen fibrils and collagen‐phosphoprotein complexes in vitro and in vivo

Mechanism of calcification: Role of collagen fibrils and collagen‐phosphoprotein complexes in vitro and in vivo

Expression and ultrastructural immunolocalization of a major 66 kDa phosphoprotein synthesized by chicken osteoblasts during mineralization in vitro

Developmental appearance and ultrastructural immunolocalization of a major 66 kDa phosphoprotein in embryonic and post‐natal chicken bone

Contact Info

Product

Resources

About