Cloning and high level expression of a synthetic gene for human basic fibroblast growth factor

Milev, Peter; Georgiev, O.I.; Tzarnoretchki, Plamen O.; Hadjiolov, A.A.

doi:10.1016/0168-1656(92)90148-3

Cited by 3 publications

(2 citation statements)

References 24 publications

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“…Aggrecan and Ng-CAM were isolated as described by Milev et al (13), and neurocan-C (the COOHterminal half of neurocan) was isolated from adult rat brain by immunoaffinity chromatography using the 1D1 monoclonal antibody (12). Recombinant human FGF-2 was prepared as described previously (14) or obtained from R & D Systems (Minneapolis, MN).…”

Section: Methodsmentioning

confidence: 99%

The Core Protein of the Chondroitin Sulfate Proteoglycan Phosphacan Is a High-affinity Ligand of Fibroblast Growth Factor-2 and Potentiates Its Mitogenic Activity

Milev

Monnerie

Popp

et al. 1998

Journal of Biological Chemistry

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Using a radioligand binding assay we have demonstrated that phosphacan, a chondroitin sulfate proteoglycan of nervous tissue that also represents the extracellular domain of a receptor-type protein tyrosine phosphatase, shows saturable, reversible, high-affinity binding (K d ϳ6 nM) to fibroblast growth factor-2 (FGF-2). Binding was reduced by only ϳ35% following chondroitinase treatment of the proteoglycan, indicating that the interaction is mediated primarily through the core protein rather than the glycosaminoglycan chains. Immunocytochemical studies also showed an overlapping localization of FGF-2 and phosphacan in the developing central nervous system. At concentrations of 10 g protein/ml, both native phosphacan and the core protein obtained by chondroitinase treatment potentiated the mitogenic effect of FGF-2 (5 ng/ml) on NIH/3T3 cells by 75-90%, which is nearly the same potentiation as that produced by heparin at an equivalent concentration. Although studies on the role of proteoglycans in mediating the binding and mitogenic effects of FGF-2 have previously focused on cell surface heparan sulfate, our results indicate that the core protein of a chondroitin sulfate proteoglycan may also regulate the access of FGF-2 to cell surface signaling receptors in nervous tissue.Phosphacan and neurocan are nervous tissue-specific chondroitin sulfate proteoglycans that are high-affinity ligands of several immunoglobulin superfamily neural cell adhesion molecules and of the extracellular matrix proteins tenascin-C and tenascin-R (1-3). These interactions are variously mediated by the chondroitin sulfate chains, N-linked oligosaccharides present on the core glycoproteins, or by other structural features of the proteoglycans that have not yet been specifically identified. Phosphacan and neurocan both bind to neurons and have potent inhibitory effects on cell adhesion and neurite outgrowth, although in certain experimental situations phosphacan may also stimulate neurite growth. Neurocan is synthesized by neurons and is a member of the family of hyaluronan-binding chondroitin sulfate proteoglycans that also includes aggrecan, versican, and brevican, whereas phosphacan, which is produced by astrocytes, is an alternative splicing product representing the extracellular domain of a receptor-type protein tyrosine phosphatase. Phosphacan is also a ligand of the neural differentiation factor HB-GAM 1 (heparin-binding growth-associated molecule)/pleiotrophin (3, 4), and both neurocan and phosphacan bind with high affinity to HB-GAM and to the related differentiation factor amphoterin, with which they colocalize in nervous tissue (3). The interactions with HB-GAM and amphoterin are largely mediated by the chondroitin sulfate chains of the proteoglycans.

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Section: Methodsmentioning

confidence: 99%

The Core Protein of the Chondroitin Sulfate Proteoglycan Phosphacan Is a High-affinity Ligand of Fibroblast Growth Factor-2 and Potentiates Its Mitogenic Activity

Milev

Monnerie

Popp

et al. 1998

Journal of Biological Chemistry

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“…A fusion histidine tag/FGF-2 protein purified from E. coli was prepared in a high yield of more than 100 mg/L of cell culture under the induction condition of 20 g of IPTG at 37°C for 5ϳ8 h. Previously, under a transcription promoter recognized by the bacteriophage T7 RNA polymerase, the yield of bFGF protein expression was approximately 40 mg/L (Squires et al 1988). Additionally, pLCII expression system by using strong PL promoter resulted in the accumulation of 20 to 25 mg of purified bFGF per liter of bacterial culture (Milev et al 1992). Compared with these processes, protein purification yield by using our production process was 3ϳ4 folds higher.…”

Section: Discussionmentioning

confidence: 86%

Optimization of treatment with recombinant FGF-2 for proliferation and differentiation of human dental stem cells, mesenchymal stem cells, and osteoblasts

Lee

Kim

Ryu

et al. 2015

Biochem. Cell Biol.

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Basic fibroblast growth factor (bFGF or FGF-2) is widely used to modulate the proliferation and differentiation of certain cell types. An expression and purification system for recombinant human FGF-2 in Escherichia coli was established for the purpose of securing a continuous supply of this protein. The purified recombinant FGF-2 significantly increased the population of human embryonic stem cells. The optimal concentrations of FGF-2 for cell proliferative induction in various adult stem cells including human dental pulp stem cells, full term human periodontal ligament stem cells, human gingival fibroblasts, mesenchymal stem cells, and osteogenic oseosarcoma were established in a dose-dependent manner. When cells were treated with recombinant FGF-2 for 6 days before osteogenic induction, the mRNA expression of the bone markers was upregulated in cells originated from human dental pulp tissue, indicating that pretreatment with FGF-2 during culture increase stem cell/progenitor population and osteogenic potential.

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Preparative two-step purification of recombinant human basic fibroblast growth factor from high-cell-density cultivation of Eschericia coli

Garke

Deckwer

Anspach

2000

Journal of Chromatography B: Biomedical Sciences and Applicatio

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Cloning and high level expression of a synthetic gene for human basic fibroblast growth factor

Cited by 3 publications

References 24 publications

The Core Protein of the Chondroitin Sulfate Proteoglycan Phosphacan Is a High-affinity Ligand of Fibroblast Growth Factor-2 and Potentiates Its Mitogenic Activity

The Core Protein of the Chondroitin Sulfate Proteoglycan Phosphacan Is a High-affinity Ligand of Fibroblast Growth Factor-2 and Potentiates Its Mitogenic Activity

Optimization of treatment with recombinant FGF-2 for proliferation and differentiation of human dental stem cells, mesenchymal stem cells, and osteoblasts

Preparative two-step purification of recombinant human basic fibroblast growth factor from high-cell-density cultivation of Eschericia coli

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