Isolation and partial molecular characterization of basic fibroblast growth factor from isolated porcine thyroid follicles and entire porcine 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-terminal-truncated 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.
Bechtner
G
G
Medizinische Klinik, Universität München, Germany.
Rieder
B
B
Linsenmaier
U
U
Kellermann
J
J
Greil
W
W
Gärtner
R
R
D000482:Allantois / Q000098:blood supply; D000595:Amino Acid Sequence; D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002823:Chorion / Q000098:blood supply; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D006493:Heparin / Q000378:metabolism; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic; D011863:Radioimmunoassay; D017386:Sequence Homology, Amino Acid; D013552:Swine; D013961:Thyroid Gland / Q000737:chemistry*
D016222:Fibroblast Growth Factor 2; D006493:Heparin