PMID: 8590594Jul 1, 1995Paper

The importance of Arg40 and 45 in the mitogenic activity and structural stability of basic fibroblast growth factor: effects of acidic amino acid substitutions

Journal of Protein Chemistry
T ArakawaG M Fox

Abstract

High-affinity binding of basic fibroblast growth factor (bFGF) to the tyrosine kinase receptor requires cell-surface heparan sulfate proteoglycan or exogenous addition of heparin. The crystal structure of bFGF shows Arg40 and 45 on the surface opposite to the heparin-binding region, suggesting that these charged residues may be involved in the receptor binding. Therefore, these amino acids were mutated to aspartic acid separately or simultaneously, and also a simultaneous mutation to glutamic acid was introduced. These mutants displayed a mitogenic activity decreased greater than tenfold compared to the wild-type protein. Addition of heparin had no effect on the activity, while these mutants showed heparin-binding characteristics resembling those of the native sequence protein. The mutants exhibited decreased stability compared to the native sequence protein. Gradual changes in conformation were observed by circular dichroic and infrared spectroscopy. Heparin chromatography also showed the presence of denatured form for these mutants. However, in the presence of multivalent anions such as citrate, sucrose octasulfate, and heparin, the conformation of the mutants resembled that of the wild-type protein, as revealed by X-ray crys...Continue Reading

References

Sep 22, 1992·Biochemistry·C R MiddaughK E Marfia
Mar 1, 1992·Archives of Biochemistry and Biophysics·S J PrestrelskiT Arakawa
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·A E ErikssonB W Matthews
Sep 1, 1991·Journal of Biochemistry·H AgoY Katsube
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·J D ZhangS R Sprang
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·K A ThomasS Fitzpatrick
Jan 23, 1987·Science·J Folkman, M Klagsbrun
Jan 1, 1987·Nouvelle Revue Française D'hématologie·A Camez, G Tobelem
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·A BairdR Guillemin
Sep 1, 1986·Journal of Cellular Physiology·D Gospodarowicz, J Cheng
Jun 30, 1986·Biochemical and Biophysical Research Communications·S UhlrichY Courtois
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·F EschR Guillemin
Nov 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·D GospodarowiczP Böhlent
Aug 30, 1983·Biochemical and Biophysical Research Communications·H Susi, D M Byler
Jan 1, 1994·Archives of Biochemistry and Biophysics·T ArakawaJ S Philo
Jan 23, 1987·Science·A T BrüngerM Karplus

❮ Previous
Next ❯

Citations

Feb 14, 1998·Nature Medicine·M ZhouT Doetschman
Dec 18, 2004·Biomaterials·Solitaire A DeLongJennifer L West
Apr 23, 2008·Current Protocols in Protein Science·T Arakawa, J Wen
Dec 24, 2002·The Journal of Biological Chemistry·Antonio FacchianoMaurizio C Capogrossi
Jan 12, 1999·Archives of Biochemistry and Biophysics·D M DanilenkoT Arakawa

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell Signaling by Tyrosine Kinases

Receptor tyrosine kinases (RTKs) are the high-affinity cell surface receptors for many polypeptide growth factors, cytokines, and hormones. RTKs have been shown not only to be key regulators of normal cellular processes but also to have a critical role in the development and progression of many types of cancer. Discover the latest research on cell signaling and RTK here.