Brain-derived fibroblast growth factor: A study of its inactivation

Westall, Fred C.; Rubin, Ruth; Gospodarowicz, Denis

doi:10.1016/0024-3205(83)90636-7

Cited by 51 publications

(33 citation statements)

References 7 publications

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“…At physiological pH and temperature, the in vitro halflifetime of bFGF activity is approximately 12 h. 23 Its binding to heparin induces a conformational change in the bFGF molecule resulting in an increased resistance against thermal denaturation and enzymatic degradation, and a reduced activation at acidic pH. 24,25 Our studies reveal that the entrapment of bFGF-heparin, 1:1 weight ratio, enabled the stabilization of bFGF within the microspheres, in agreement with the stoichiometry of heparin binding to bFGF.…”

Section: Discussionsupporting

confidence: 70%

Enhancing the vascularization of three‐dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres

Perets

Baruch

Weisbuch

et al. 2003

J Biomedical Materials Res

416

270

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Copyright: Ren et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT As a promising strategy for the successful regeneration of articular cartilage, tissue engineering has received increasing recognition of control release. Two kinds of functional poly (alanine ethyl ester-co-glycine ethyl ester) phosphazene microspheres with different ratios of side-substituent groups were synthesized by emulsion technique. The rate of degradation/hydrolysis of the polymers was carefully tuned to suit the desired application for control release. For controlled delivery of growth factors, the microspheres overcame most of severe side effects linked to demineralized bone matrix (DBM) scaffolds, which had been previously optimized for cartilage regeneration. The application of scaffolds in chondrogenic differentiation was investigated by subcutaneous implantation in nude mice. In the present study, we have provided a novel microsphere-incorporating demineralized bone matrix (MS/DBM) scaffolds to release transforming growth factor-β1 or insulin-like growth factors-1. Laser confocal fluorescence staining showed that the surface of microspheres was a suitable environment for cell attachment. Histological and immunohistochemical evaluations have shown that significantly more cartilaginous extracellular matrix was detected in MS/DBM group when compared with DBM alone group (P<0.05). In addition, the biomechanical test showed that this composite scaffold exhibited favorable mechanical strength as a delivery platform. In conclusion, we demonstrated that MS/DBM scaffolds was sufficient to support stem bone marrow-derived mesenchymal stem cells chondrogenesis and neo-cartilage formation.

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Section: Discussionsupporting

confidence: 70%

Enhancing the vascularization of three‐dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres

Perets

Baruch

Weisbuch

et al. 2003

J Biomedical Materials Res

416

270

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“…In previous studies it has been shown that even at 37°C and at neutral pHs, the half-life of FGF is 24 h (Westall et al, 1983). Interaction of heparin with FGF will likely result in greatly extending its half-life.…”

Section: Heparin and Hhs-4 Reduced The Ability Of Egfmentioning

confidence: 98%

Heparin protects basic and acidic FGF from inactivation

Gospodarowicz

Cheng

1986

Journal Cellular Physiology

825

406

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The ability of heparin or that of hexuronyl hexosaminoglycan sulfate (HHS-4) to protect basic or acidic fibroblast growth factor (FGF) from acid or heat inactivation has been analyzed. Both freshly prepared basic and acidic FGF stimulate the growth of baby hamster kidney (BHK-21) cells exposed to medium supplemented with transferrin and insulin. Freshly prepared basic FGF was 10 fold more potent than acidic FGF. The addition of heparin to the medium decreased the potency of basic FGF while it potentiated that of acidic FGF. Upon storage of FGF at -80 degrees C, a decline in potency of both basic and acidic FGF was observed. Heparin, when added to the medium, potentiated their activities, which became similar to that of freshly prepared basic FGF. In order to test whether heparin could protect basic or acidic FGF from inactivation, both mitogens were exposed to acid conditions (1% trifluoroacetic acid, pH 1.08, 2 h) or heat (65 degrees C, 5 min) which inactivate basic or acidic FGF. When exposed to such treatment in the presence of heparin or HHS-4, basic and acidic FGF retained their potency. The effect of heparin and HHS-4 on the bioactivity of basic and acidic FGF is truly of a protective nature, since they had no effect when added after inactivation of the mitogens. Potentiation of the bioactivity of the protected mitogens or of the inactivated one could only be observed when cells were exposed to high heparin or HHS-4 concentrations. This indicates that heparin and HHS-4, in addition to protecting FGF from inactivation, also acts at another locus, as yet unidentified.

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“…Residues Lys 128 , Arg 129 , Lys 134 , and Lys 138 * have been identified as part of the heparin-binding domain of FGF-2 (Li et al, 1994;Thompson et al, 1994). Heparin has also been shown to protect FGF-2 from inactivation resulting from exposure to low pH, elevated temperature or proteases, and to restore bioactivity to inactive growth factor (Westall et al, 1983;Gospodarowicz and Cheng, 1986;Saksela et al, 1988;Sommer and Rifkin, 1989;Pineda-Lucena et al, 1994a).…”

Section: Introductionmentioning

confidence: 99%

1H, 15N, 13C and 13CO assignments and secondary structure determination of basic fibroblast growth factor using 3D heteronuclear NMR spectroscopy

et al. 1995

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Supplementary material-tables containing the 1 H, 15 N, 13 CO, and 13 C resonance assignments for the major and minor conformations of FGF-2-is available in the Biological Magnetic Resonance Data Bank, BMRB 4091, at http://www.bmrb.wisc.edu/data_library/summary/index.php?bmrbId=4091. AbstractThe assignments of the 1 H, 15 N, 13 CO, and 13 C resonances of recombinant human basic fibroblast growth factor (FGF-2), a protein comprising 154 residues and with a molecular mass of 17.2 kDa, is presented based on a series of three-dimensional triple-resonance heteronuclear NMR experiments. These studies employ uniformly labeled 15 N-and 15 N-/ 13 C-labeled FGF-2 with an isotope incorporation > 95% for the protein expressed in E. coli. The sequence-specific backbone assignments were based primarily on the interresidue correlation of C α , C β , and H α to the backbone amide 1 H and 15 N of the next residue in the CBCA(CO)NH and HBHA(CO)NH experiments and the intraresidue correlation of C α , C β , and H α to the backbone amide 1 H and 15 N in the CBCANH and HNHA experiments. In addition, C α and C β chemical shift assignments were used to determine amino acid types. Sequential assignments were verified from carbonyl correlations observed in the HNCO and2 HCACO experiments and C α correlations from the HNCA experiment. Aliphatic side-chain spin systems were assigned primarily from H(CCO)NH and C(CO)NH experiments that correlate all the aliphatic 1 H and 13 C resonances of a given residue with the amide resonance of the next residue. Additional side-chain assignments were made from HCCH-COSY and HCCH-TOCSY experiments. The secondary structure of FGF-2 is based on NOE data involving the NH, H α , and H β protons as well as 3 JH N H α coupling constants, amide exchange, and 13 C α and 13 C β secondary chemical shifts. It is shown that FGF-2 consists of 11 well-defined antiparallel β-sheets (residues 30-34, 39-44, 48-53, 62-67, 71-76, 81-85, 91-94, 103-108, 113-118, 123-125, and 148-152) and a helix-like structure (residues 131-136), which are connected primarily by tight turns. This structure differs from the refined X-ray crystal structures of FGF-2, where residues 131-136 were defined as β-strand XI. The discovery of the helix-like region in the primary heparin-binding site (residues 128-138) instead of the β-strand conformation described in the X-ray structures may have important implications in understanding the nature of heparin-FGF-2 interactions. In addition, two distinct conformations exist in solution for the N-terminal residues 9-28. This is consistent with the X-ray structures of FGF-2, where the first 17-19 residues were ill defined.

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Brain-derived fibroblast growth factor: A study of its inactivation

Cited by 51 publications

References 7 publications

Enhancing the vascularization of three‐dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres

Enhancing the vascularization of three‐dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres

Heparin protects basic and acidic FGF from inactivation

1H, 15N, 13C and 13CO assignments and secondary structure determination of basic fibroblast growth factor using 3D heteronuclear NMR spectroscopy

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