Isolation and partial molecular characterization of basic fibroblast growth factor from isolated porcine thyroid follicles and entire porcine thyroid glands

1993 ◽  
Vol 129 (5) ◽  
pp. 458-466 ◽  
Author(s):  
Günther Bechtner ◽  
Bernhard Rieder ◽  
Ullrich Linsenmaier ◽  
J Kellermann ◽  
Walter Greil ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Sodium Dodecyl ◽  
Fibroblast Growth ◽  
Thyroid Cells ◽  
Amino Terminal ◽  
Thyroid Glands

Immunoreactive basic fibroblast growth factor (bFGF) could be isolated from the cytosol preparation of isolated porcine thyroid follicles as well as in the conditioned medium from thyroid follicles in suspension culture. A double band with 16 500 and 15 500 D was detected on sodium dodecyl sulfate polyacrylamide gel electrophoresis. In dot blot and western blot the isolated peptide was immunoreactive with a specific anti-bovine bFGF antibody. For further biochemical characterization, bFGF was isolated from entire porcine thyroid glands by ammonium sulfate precipitation, cation exchange chromatography and heparin affinity chromatography. The material obtained from all three origins was identical concerning affinity to heparin and immunoreactivity with the specific anti-bovine bFGF antibody and induced neovascularization in the chorioallantois membranes of chick embryos. Amino acid sequence analysis of the 16-amino-terminal amino acids of the isolated bFGF was in accordance with the established complete 146-amino-acid bFGF molecule except that glycine in position 10 is replaced by phenylalanine. An additionally identified minor peptide presumably is an amino-terminaltruncated form of bFGF, missing the first 15 amino acids. We conclude that the physiological significance of bFGF released by thyroid cells may be the regulation of angiogenesis during thyroid development and goiter growth.

scholarly journals Nuclear Activities of Basic Fibroblast Growth Factor: Potentiation of Low-Serum Growth Mediated by Natural or Chimeric Nuclear Localization Signals

1999 ◽  
Vol 10 (5) ◽  
pp. 1429-1444 ◽  
Author(s):  
Marco Arese ◽  
Yan Chen ◽  
Robert Z. Florkiewicz ◽  
Anna Gualandris ◽  
Bin Shen ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Serum Starvation ◽  
Fibroblast Growth ◽  
Stable Transfectants ◽  
Nuclear Localization Signals ◽  
Amino Terminal ◽  
Low Serum

Human basic fibroblast growth factor (FGF-2) occurs in four isoforms: a low molecular weight (LMW FGF-2, 18 kDa) and three high molecular weight (HMW FGF-2, 22, 22.5, and 24 kDa) forms. LMW FGF-2 is primarily cytoplasmic and functions in an autocrine manner, whereas HMW FGF-2s are nuclear and exert activities through an intracrine, perhaps nuclear, pathway. Selective overexpression of HMW FGF-2 forms in fibroblasts promotes growth in low serum, whereas overexpression of LMW FGF-2 does not. The HMW FGF-2 forms have two functional domains: an amino-terminal extension and a common 18-kDa amino acid sequence. To investigate the role of these regions in the intracrine signaling of HMW FGF-2, we produced stable transfectants of NIH 3T3 fibroblasts overexpressing either individual HMW FGF-2 forms or artificially nuclear-targeted LMW FGF-2. All of these forms of FGF-2 localize to the nucleus/nucleolus and induce growth in low serum. The nuclear forms of FGF-2 trigger a mitogenic stimulus under serum starvation conditions and do not specifically protect the cells from apoptosis. These data indicate the existence of a specific role for nuclear FGF-2 and suggest that LMW FGF-2 represents the biological messenger in both the autocrine/paracrine and intracrine FGF-2 pathways.

Purification and characterization of the 210-amino acid recombinant basic fibroblast growth factor form (FGF-2)

FEBS Letters ◽  
1994 ◽  
Vol 349 (1) ◽  
pp. 23-28 ◽  
Author(s):  
Véronique Patry ◽  
Béatrix Bugler ◽  
François Amalric ◽  
Jean-Claude Promé ◽  
Hervé Prats
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Fibroblast Growth ◽  
Purification And Characterization ◽  
Factor Form

scholarly journals Endogenous basic fibroblast growth factor isoforms involved in different intracellular protein complexes

1997 ◽  
Vol 326 (1) ◽  
pp. 259-264 ◽  
Author(s):  
Véronique PATRY ◽  
Béatrix BUGLER ◽  
Arlette MARET ◽  
Michel POTIER ◽  
Hervé PRATS
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Cho Cells ◽  
Protein Complexes ◽  
Biological Activities ◽  
Molecular Complexes ◽  
Fibroblast Growth ◽  
Initiation Of Translation

Four forms of basic fibroblast growth factor (bFGF or FGF-2) result from an alternative initiation of translation involving one AUG (155-amino acid form) and three CUGs (210-, 201- and 196-amino acid forms). These different forms of bFGF show different intracellular biological activities. To identify their intracellular targets, the 210- and 155-amino acid forms of bFGF were independently transfected into CHO cells and their correct subcellular localizations were verified, the 155-amino acid bFGF form being essentially cytoplasmic whereas the 210-amino acid protein was nuclear. The radiation fragmentation method was used to determine the target size of the different bFGF isoforms in the transfected CHO cells and to show that the 210- and 155-amino acids bFGF isoforms were included in protein complexes of 320 and 130 kDa respectively. Similar results were obtained using the SK-Hep1 cell line, which naturally expressed all forms of bFGF. Co-immunoprecipitation assays using different chimaeric bFGF–chloramphenicol acetyltransferase proteins showed that different cellular proteins are associated with different parts of the bFGF molecule. We conclude that bFGF isoforms are involved in different molecular complexes in the cytosol and nucleus, which would reflect different functions for these proteins.

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