PMID: 2106873Feb 15, 1990Paper

Isolation and characterization of proteoglycans synthesized by mouse osteoblastic cells in culture during the mineralization process

The Biochemical Journal
Y TakeuchiY Shishiba

Abstract

Proteoglycans in mineralized (0.5 M-EDTA/4 M-guanidinium chloride-extractable) and non-mineralized (4 M-guanidinium chloride-extractable) matrices synthesized by a mouse osteoblastic-cell line MC3T3-E1 were characterized at different phases of mineralization in vitro. Cell cultures were labelled with [35S]sulphate and either [3H]glucosamine or 3H-labelled amino acids. At the mineralization phase a large majority of proteoglycans were extracted with 4 M-guanidinium chloride (G extract), and at least five species of labelled proteoglycans were identified; dermatan sulphate proteoglycans (DSPG), apparent Mr approx. 120,000 and 70,000), heparan sulphate proteoglycans (HSPG, apparent Mr approx. 200,000 and 120,000) and DS chains with very little core protein. DSPGs weakly bound to an octyl-Sepharose CL-4B column and HSPGs bound more tightly, whereas DS chains did not bind to the column. Amounts of labelled proteoglycans extracted with 0.5 M-EDTA/4 M-guanidinium chloride (EDTA extract) were much less than those in G extract. Although the predominant species in the EDTA extract were comparable with the DS or DSPGs in the G extract, none of them bound to octyl-Sepharose CL-4B, indicating their lack of hydrophobicity. At the nonminerali...Continue Reading

Citations

Jan 1, 1996·Connective Tissue Research·A L Boskey
Feb 5, 1999·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·K HoshiH Ozawa
Feb 24, 2001·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·K HoshiH Ozawa
Oct 27, 2015·Journal of Structural Biology·A BrionM Rousseau
Aug 23, 2008·Journal of Cellular Physiology·Tatsuya MiyazakiOsamu Suzuki
Mar 8, 2005·Matrix Biology : Journal of the International Society for Matrix Biology·Dealba GheduzziIvonne Pasquali Ronchetti
Jul 10, 2002·Archives of Oral Biology·S YonedaM Yanagishita
Feb 1, 1994·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·M SlaterR S Mason
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Jul 1, 1993·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Y TakeuchiY Shishiba
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Oct 23, 1997·Biochemical and Biophysical Research Communications·T KobayashiK Chihara
Apr 2, 2002·Biochemical and Biophysical Research Communications·Sarah G ReesGraham Embery

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