Stabilizing basic fibroblast growth factor using protein engineering

Masaharu Seno, Reiko Sasada, Makoto Iwane, Katsuichi Sudo, Tsutomu Kurokawa, Kumiko Ito, Koichi Igarashi

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

Using site directed mutagenesis, each of the four cysteines present at amino acid residues 26, 70, 88, and 93 of the mature protein of human basic fibroblast growth factor (bFGF) was individually changed to serine. The biological activity and heparin binding ability was retained when the serine was substituted for the cysteine residue at either 70 or 88 of the bFGF protein. This finding indicates that the cysteines at these positions are not essential for expressing biological activity. The substitution of the residues at these positions, especially at position 88, reduced the heterogeneity recognized as several peaks of bFGF eluted from a heparin affinity column, even after oxidation with hydrogen peroxide, suggesting that the cysteines at these positions are exposed to the surface of the molecule to form disulfide bonds that induce heterologous conformations. Furthermore, under acidic conditions, these modified bFGFs are revealed to be more stable in maintaining their activity. These facts suggest that this protein has been successfully modified by protein engineering.

Original languageEnglish
Pages (from-to)701-708
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume151
Issue number2
DOIs
Publication statusPublished - Mar 15 1988
Externally publishedYes

Fingerprint

Protein Engineering
Fibroblast Growth Factor 2
Cysteine
Bioactivity
Serine
Heparin
Proteins
Mutagenesis
Site-Directed Mutagenesis
Disulfides
Hydrogen Peroxide
Conformations
Substitution reactions
Amino Acids
Oxidation
Molecules

Keywords

  • basic fibroblast growth factor
  • bFGF,
  • bovine serum albumin
  • BSA,
  • cDNA,
  • complementary DNA
  • FGF,
  • fibroblast growth factor
  • sodium dodecyl sulfate
  • SOS,

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Stabilizing basic fibroblast growth factor using protein engineering. / Seno, Masaharu; Sasada, Reiko; Iwane, Makoto; Sudo, Katsuichi; Kurokawa, Tsutomu; Ito, Kumiko; Igarashi, Koichi.

In: Biochemical and Biophysical Research Communications, Vol. 151, No. 2, 15.03.1988, p. 701-708.

Research output: Contribution to journalArticle

Seno, Masaharu ; Sasada, Reiko ; Iwane, Makoto ; Sudo, Katsuichi ; Kurokawa, Tsutomu ; Ito, Kumiko ; Igarashi, Koichi. / Stabilizing basic fibroblast growth factor using protein engineering. In: Biochemical and Biophysical Research Communications. 1988 ; Vol. 151, No. 2. pp. 701-708.
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AU - Igarashi, Koichi

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