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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Jan;81(2):357–361. doi: 10.1073/pnas.81.2.357

Purification and characterization of acidic fibroblast growth factor from bovine brain.

K A Thomas, M Rios-Candelore, S Fitzpatrick
PMCID: PMC344675  PMID: 6582493

Abstract

Acidic brain fibroblast growth factor has been purified a minimum of 35,000-fold to apparent homogeneity by a combination of differential salt precipitation, ion exchange chromatography, gel filtration, isoelectric focusing, and hydrophobic chromatography on a C4 reversed-phase HPLC column. Two microheterogeneous forms of the molecule are obtained with apparent molecular masses of 16,600 and 16,800 daltons. The mitogen is highly active with half-maximal stimulation of BALB/c 3T3 fibroblasts at about 40 pg/ml in an assay using incorporation of [methyl-3H]thymidine into DNA.

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Selected References

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  1. Antoniades H. N. Human platelet-derived growth factor (PDGF): purification of PDGF-I and PDGF-II and separation of their reduced subunits. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7314–7317. doi: 10.1073/pnas.78.12.7314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anzano M. A., Roberts A. B., Smith J. M., Lamb L. C., Sporn M. B. Purification by reverse-phase high-performance liquid chromatography of an epidermal growth factor-dependent transforming growth factor. Anal Biochem. 1982 Sep 1;125(1):217–224. doi: 10.1016/0003-2697(82)90405-5. [DOI] [PubMed] [Google Scholar]
  3. Assoian R. K., Komoriya A., Meyers C. A., Miller D. M., Sporn M. B. Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem. 1983 Jun 10;258(11):7155–7160. [PubMed] [Google Scholar]
  4. Barritault D., Plouët J., Courty J., Courtois Y. Purification, characterization, and biological properties of the eye-derived growth factor from retina: analogies with brain-derived growth factor. J Neurosci Res. 1982;8(2-3):477–490. doi: 10.1002/jnr.490080235. [DOI] [PubMed] [Google Scholar]
  5. Binoux M., Hossenlopp P., Lassarre C., Hardouin N. Production of insulin-like growth factors and their carrier by rat pituitary gland and brain explants in culture. FEBS Lett. 1981 Feb 23;124(2):178–184. doi: 10.1016/0014-5793(81)80131-7. [DOI] [PubMed] [Google Scholar]
  6. Deuel T. F., Huang J. S., Proffitt R. T., Baenziger J. U., Chang D., Kennedy B. B. Human platelet-derived growth factor. Purification and resolution into two active protein fractions. J Biol Chem. 1981 Sep 10;256(17):8896–8899. [PubMed] [Google Scholar]
  7. Gambarini A. G., Armelin H. A. Purification and partial characterization of an acidic fibroblast growth factor from bovine pituitary. J Biol Chem. 1982 Aug 25;257(16):9692–9697. [PubMed] [Google Scholar]
  8. Gospodarowicz D., Bialecki H., Greenburg G. Purification of the fibroblast growth factor activity from bovine brain. J Biol Chem. 1978 May 25;253(10):3736–3743. [PubMed] [Google Scholar]
  9. Gospodarowicz D., Lui G. M., Cheng J. Purification in high yield of brain fibroblast growth factor by preparative isoelectric focusing at pH 9.6. J Biol Chem. 1982 Oct 25;257(20):12266–12276. [PubMed] [Google Scholar]
  10. Heldin C. H., Westermark B., Wasteson A. Platelet-derived growth factor: purification and partial characterization. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3722–3726. doi: 10.1073/pnas.76.8.3722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Illingworth J. A. A common source of error in pH measurements. Biochem J. 1981 Apr 1;195(1):259–262. doi: 10.1042/bj1950259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kellett J. G., Tanaka T., Rowe J. M., Shiu R. P., Friesen H. G. The characterization of growth factor activity in human brain. J Biol Chem. 1981 Jan 10;256(1):54–58. [PubMed] [Google Scholar]
  13. Lemmon S. K., Bradshaw R. A. Purification and partial characterization of bovine pituitary fibroblast growth factor. J Cell Biochem. 1983;21(3):195–208. doi: 10.1002/jcb.240210302. [DOI] [PubMed] [Google Scholar]
  14. Lemmon S. K., Riley M. C., Thomas K. A., Hoover G. A., Maciag T., Bradshaw R. A. Bovine fibroblast growth factor: comparison of brain and pituitary preparations. J Cell Biol. 1982 Oct;95(1):162–169. doi: 10.1083/jcb.95.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maciag T., Hoover G. A., Weinstein R. High and low molecular weight forms of endothelial cell growth factor. J Biol Chem. 1982 May 25;257(10):5333–5336. [PubMed] [Google Scholar]
  16. Marquardt H., Todaro G. J. Human transforming growth factor. Production by a melanoma cell line, purification, and initial characterization. J Biol Chem. 1982 May 10;257(9):5220–5225. [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Rinderknecht E., Humbel R. E. Polypeptides with nonsuppressible insulin-like and cell-growth promoting activities in human serum: isolation, chemical characterization, and some biological properties of forms I and II. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2365–2369. doi: 10.1073/pnas.73.7.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rinderknecht E., Humbel R. E. Primary structure of human insulin-like growth factor II. FEBS Lett. 1978 May 15;89(2):283–286. doi: 10.1016/0014-5793(78)80237-3. [DOI] [PubMed] [Google Scholar]
  20. Rinderknecht E., Humbel R. E. The amino acid sequence of human insulin-like growth factor I and its structural homology with proinsulin. J Biol Chem. 1978 Apr 25;253(8):2769–2776. [PubMed] [Google Scholar]
  21. Rubin J. S., Mariz I., Jacobs J. W., Daughaday W. H., Bradshaw R. A. Isolation and partial sequence analysis of rat basic somatomedin. Endocrinology. 1982 Mar;110(3):734–740. doi: 10.1210/endo-110-3-734. [DOI] [PubMed] [Google Scholar]
  22. Simpson R. J., Neuberger M. R., Liu T. Y. Complete amino acid analysis of proteins from a single hydrolysate. J Biol Chem. 1976 Apr 10;251(7):1936–1940. [PubMed] [Google Scholar]
  23. Taylor J. M., Mitchell W. M., Cohen S. Epidermal growth factor. Physical and chemical properties. J Biol Chem. 1972 Sep 25;247(18):5928–5934. [PubMed] [Google Scholar]
  24. Thomas K. A., Riley M. C., Lemmon S. K., Baglan N. C., Bradshaw R. A. Brain fibroblast growth factor: nonidentity with myelin basic protein fragments. J Biol Chem. 1980 Jun 25;255(12):5517–5520. [PubMed] [Google Scholar]
  25. Westall F. C., Lennon V. A., Gospodarowicz D. Brain-derived fibroblast growth factor: identity with a fragment of the basic protein of myelin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4675–4678. doi: 10.1073/pnas.75.10.4675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]

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