Molecular characterization of recombinant human acidic fibroblast growth factor produced in E. coli: Comparative studies with human basic fibroblast growth factor

Tatsuya Watanabe, Masaharu Seno, Reiko Sasada, Koichi Igarashi

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

Synthetic cDNA coding for human acidic fibroblast growth factor (haFGF) was expressed in E. coli under the control of the T7 promoter. The haFGF produced was purified extensively using heparin-Sepharose and phenyl-Sepharose columns. The mitogenic activity of haFGF on 3T3 and endothelial cells was significantly potentiated in the presence of heparin (10-50 μg|ml), while angiogenic activity was observed on chick embryo chorioallantoic membrane without exogenously added heparin. This significant potentiation of mitogenic activity was observed specifically with haFGF, not human basic fibroblast growth factor (hbFGF). Circular dichroism spectra of haFGF was not affected by the presence of heparin. The affinity of haFGF for heparin was examined using heparin affinity HPLC and was precisely confirmed to be relatively lower than that of hbFGF. These results implied that haFGF was potentiated by heparin and that this potentiation did not involve a significant change in the conformation of the haFGF molecule. The affinity of haFGF for copper was also confirmed to be higher than that of hbFGF using a copper affinity HPLC column. In addition, under acidic conditions, haFGF appeared more stable than hbFGF and was further stabilized in the presence of heparin.

Original languageEnglish
Pages (from-to)869-879
Number of pages11
JournalMolecular Endocrinology
Volume4
Issue number6
Publication statusPublished - Jun 1990
Externally publishedYes

Fingerprint

Fibroblast Growth Factor 1
Fibroblast Growth Factor 2
Escherichia coli
Heparin
Copper
High Pressure Liquid Chromatography
Chorioallantoic Membrane
3T3 Cells
Chick Embryo
Circular Dichroism
Human Activities

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Molecular characterization of recombinant human acidic fibroblast growth factor produced in E. coli : Comparative studies with human basic fibroblast growth factor. / Watanabe, Tatsuya; Seno, Masaharu; Sasada, Reiko; Igarashi, Koichi.

In: Molecular Endocrinology, Vol. 4, No. 6, 06.1990, p. 869-879.

Research output: Contribution to journalArticle

@article{7699b648f8f24f7fae63a505edb9572b,
title = "Molecular characterization of recombinant human acidic fibroblast growth factor produced in E. coli: Comparative studies with human basic fibroblast growth factor",
abstract = "Synthetic cDNA coding for human acidic fibroblast growth factor (haFGF) was expressed in E. coli under the control of the T7 promoter. The haFGF produced was purified extensively using heparin-Sepharose and phenyl-Sepharose columns. The mitogenic activity of haFGF on 3T3 and endothelial cells was significantly potentiated in the presence of heparin (10-50 μg|ml), while angiogenic activity was observed on chick embryo chorioallantoic membrane without exogenously added heparin. This significant potentiation of mitogenic activity was observed specifically with haFGF, not human basic fibroblast growth factor (hbFGF). Circular dichroism spectra of haFGF was not affected by the presence of heparin. The affinity of haFGF for heparin was examined using heparin affinity HPLC and was precisely confirmed to be relatively lower than that of hbFGF. These results implied that haFGF was potentiated by heparin and that this potentiation did not involve a significant change in the conformation of the haFGF molecule. The affinity of haFGF for copper was also confirmed to be higher than that of hbFGF using a copper affinity HPLC column. In addition, under acidic conditions, haFGF appeared more stable than hbFGF and was further stabilized in the presence of heparin.",
author = "Tatsuya Watanabe and Masaharu Seno and Reiko Sasada and Koichi Igarashi",
year = "1990",
month = "6",
language = "English",
volume = "4",
pages = "869--879",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "6",

}

TY - JOUR

T1 - Molecular characterization of recombinant human acidic fibroblast growth factor produced in E. coli

T2 - Comparative studies with human basic fibroblast growth factor

AU - Watanabe, Tatsuya

AU - Seno, Masaharu

AU - Sasada, Reiko

AU - Igarashi, Koichi

PY - 1990/6

Y1 - 1990/6

N2 - Synthetic cDNA coding for human acidic fibroblast growth factor (haFGF) was expressed in E. coli under the control of the T7 promoter. The haFGF produced was purified extensively using heparin-Sepharose and phenyl-Sepharose columns. The mitogenic activity of haFGF on 3T3 and endothelial cells was significantly potentiated in the presence of heparin (10-50 μg|ml), while angiogenic activity was observed on chick embryo chorioallantoic membrane without exogenously added heparin. This significant potentiation of mitogenic activity was observed specifically with haFGF, not human basic fibroblast growth factor (hbFGF). Circular dichroism spectra of haFGF was not affected by the presence of heparin. The affinity of haFGF for heparin was examined using heparin affinity HPLC and was precisely confirmed to be relatively lower than that of hbFGF. These results implied that haFGF was potentiated by heparin and that this potentiation did not involve a significant change in the conformation of the haFGF molecule. The affinity of haFGF for copper was also confirmed to be higher than that of hbFGF using a copper affinity HPLC column. In addition, under acidic conditions, haFGF appeared more stable than hbFGF and was further stabilized in the presence of heparin.

AB - Synthetic cDNA coding for human acidic fibroblast growth factor (haFGF) was expressed in E. coli under the control of the T7 promoter. The haFGF produced was purified extensively using heparin-Sepharose and phenyl-Sepharose columns. The mitogenic activity of haFGF on 3T3 and endothelial cells was significantly potentiated in the presence of heparin (10-50 μg|ml), while angiogenic activity was observed on chick embryo chorioallantoic membrane without exogenously added heparin. This significant potentiation of mitogenic activity was observed specifically with haFGF, not human basic fibroblast growth factor (hbFGF). Circular dichroism spectra of haFGF was not affected by the presence of heparin. The affinity of haFGF for heparin was examined using heparin affinity HPLC and was precisely confirmed to be relatively lower than that of hbFGF. These results implied that haFGF was potentiated by heparin and that this potentiation did not involve a significant change in the conformation of the haFGF molecule. The affinity of haFGF for copper was also confirmed to be higher than that of hbFGF using a copper affinity HPLC column. In addition, under acidic conditions, haFGF appeared more stable than hbFGF and was further stabilized in the presence of heparin.

UR - http://www.scopus.com/inward/record.url?scp=0025155546&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025155546&partnerID=8YFLogxK

M3 - Article

C2 - 1700280

AN - SCOPUS:0025155546

VL - 4

SP - 869

EP - 879

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 6

ER -