Characterization of a cysteine-free analog of recombinant human basic fibroblast growth factor

Biochemical and Biophysical Research Communications
T ArakawaG M Fox

Abstract

Using oligo site-directed mutagenesis, we have modified our synthetic gene for human basic fibroblast growth factor (bFGF) to replace all four cysteine codons with serine codons. The corresponding protein was expressed in Escherichia coli and purified from inclusion bodies by solubilization in urea followed by a series of column chromatographies and a folding step. The resulting protein, having no cysteine residues, is unable to form either intramolecular or intermolecular disulfide bonds. The secondary and tertiary structures of the purified analog, as determined by circular dichroism and fluorescence spectroscopy, were identical within experimental error to recombinant bovine and human bFGF with unaltered amino acid sequences. Reflecting the similar conformation, the analog protein exhibited mitogenic activity on NIH 3T3 cells which was indistinguishable from the natural sequence molecule.

References

Mar 15, 1988·Biochemical and Biophysical Research Communications·M SenoK Igarashi
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·A BairdR Guillemin

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Citations

Aug 1, 1991·Journal of Protein Chemistry·C S WuJ T Yang
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·A E ErikssonB W Matthews
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·J D ZhangS R Sprang
Oct 15, 1996·European Journal of Biochemistry·A RomeroG Giménez-Gallego
Jan 1, 1991·Annals of the New York Academy of Sciences·A SeddonP Böhlen
Mar 30, 2004·Biochemical and Biophysical Research Communications·Yutaka YamamotoNaomi Kondo
Aug 15, 1991·Archives of Biochemistry and Biophysics·R A CopelandD Marquis-Omer
Sep 12, 1997·The Journal of Biological Chemistry·H EmotoN Itoh
Jan 12, 1999·Archives of Biochemistry and Biophysics·D M DanilenkoT Arakawa
Feb 15, 1991·Archives of Biochemistry and Biophysics·S KotakeR A Copeland
Nov 1, 1992·Archives of Biochemistry and Biophysics·R K DukorT Arakawa
Mar 1, 1992·Archives of Biochemistry and Biophysics·S J PrestrelskiT Arakawa

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