Isolation of an amino terminal extended form of basic fibroblast growth factor |
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| Authors: | N Ueno A Baird F Esch N Ling R Guillemin |
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| Institution: | 1. Cell-Biomaterial Biohybrid Systems, Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, 2500 boul. de l''Université, Sherbrooke, Québec J1K 2R1, Canada;2. Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, 2500 boul. de l''Université, Sherbrooke, Québec J1K 2R1, Canada;1. Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, 2500 boul. de l''Université, Sherbrooke, Québec J1K 2R1, Canada;2. Clinical Research Center of Centre Hospitalier Universitaire de Sherbrooke, 12e Avenue N, Sherbrooke, Québec J1H 5N4, Canada;3. Pharmacology Institute of Sherbrooke, 12e Avenue N, Sherbrooke, Québec J1H 5N4, Canada;1. Wise Laboratory of Environmental and Genetic Toxicology, Department of Pharmacology and Toxicology, University of Louisville, 505 S. Prescott Street, Louisville, KY 40292, USA;2. Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, Science Building, 96 Falmouth Street, Portland, ME 04103, USA;3. Wise Laboratory Field Research Program, 1320 19th Street, NW, 5th Floor, Washington, DC 20036, USA;4. Maine Center for Toxicology and Environmental Health, University of Southern Maine, Science Building, 96 Falmouth Street, Portland, ME 04103, USA;5. Department of Applied Medical Science, University of Southern Maine, Science Building, 96 Falmouth Street, Portland, ME 04103, USA;6. Exponent, Inc., 1800 Diagonal Road, Suite 500, Alexandria, VA 22314, USA;7. Center for Environmental Sciences and Engineering, University of Connecticut, 3107 Horsebarn Hill Road, U-4210, Storrs, CT 06269, USA;8. Yale School of Public Health, P.O. Box 208034, 60 College Street, New Haven, CT 06520, USA |
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| Abstract: | Extraction of bovine pituitaries in the presence of enzyme inhibitors (2 mM PMSF, 2 mM sodium tetrathionate, 15 microM pepstatin A, and 1 mM EDTA) resulted in the isolation of two distinct forms of basic fibroblast growth factor. Partial characterization of both molecules showed one form to be identical to basic FGF(1-146) which has already been reported by our laboratory. The second form was estimated by SDS-PAGE to have a molecular weight of 17,000 Daltons which is slightly larger than that of basic FGF(1-146). Amino acid analysis shows the presence of 8 new residues more than basic FGF(1-146) which accounts for the difference in molecular weight. Gas-phase sequencing of this molecule indicated that it bears a blocked amino terminus. Furthermore, this higher molecular weight form of basic FGF did not show immunoreactivity with antibodies specific for the amino terminus of basic FGF(1-146) but cross reacted with antibodies generated against midportion fragments of basic FGF(1-146), indicating that the molecule is amino terminally extended. Like basic FGF(1-146), the molecule is a potent mitogenic factor for vascular endothelial cells. Taken together these results demonstrate the existence of a precursor form of basic FGF which is extended by 8 residues at the amino terminus with the first residue being blocked. |
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