Structure-function studies of heparin-binding (acidic fibroblast) growth factor-1 using site-directed mutagenesis.
The heparin-binding or fibroblast growth factors (HBGFs) modulate cell growth and migration, angiogenesis, wound repair, neurite extension, and mesoderm induction. Relatively little is known regarding the precise mechanism of action of these growth factors or the structural basis for their action. A better understanding of the structural basis for the different activities of these proteins should lead to the development of agonists and antagonists of specific HBGF activities. In this report, we summarize evidence that indicates that the heparin-binding and mitogenic activities of HBGF-1 can be dissociated from the receptor-binding activities of the growth factor by site-directed mutagenesis of a single lysine residue. Thus, the mutant HBGF-1 has normal receptor-binding activity and is capable of stimulating tyrosine kinase activity and proto-oncogene expression but is not able to elicit a mitogenic response. A similar dissociation of early events such as proto-oncogene expression from the mitogenic response is observed when the human wild-tupe HBGF-1 is used in the absence of added heparin. These results indicate that intracellular sites of action by the growth factor may be required to complete the mitogenic response. Further evidence for this idea is provided by transfection experiments where NIH 3T3 cells are engineered to produce large quantities of wild-type or mutant HBGF-1. Production of wild-type induces a transformed phenotype, whereas over-production of the mutant does not. The majority of both forms of the protein is found in the nuclear fraction of the transfected cells. Additional site-directed mutagenesis of putative nuclear translocation sequences in the wild-type protein do not affect mitogenic activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Burgess
W H
WH
American Red Cross, Jerome H. Holland Laboratory for the Biomedical Sciences, Rockville, Maryland 20855.
Shaheen
A M
AM
Hampton
B
B
Donohue
P J
PJ
Winkles
J A
JA
D000818:Animals; D002478:Cells, Cultured; D004790:Enzyme Induction / Q000187:drug effects; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects; D006493:Heparin / Q000378:metabolism; D008239:Lysine; D051379:Mice; D008934:Mitogens / Q000235:genetics / Q000494:pharmacology; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011487:Protein Conformation; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000494:pharmacology; D013329:Structure-Activity Relationship
C000715470:MAS1 protein, human; D008934:Mitogens; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D011993:Recombinant Fusion Proteins; D016220:Fibroblast Growth Factor 1; D006493:Heparin; D008239:Lysine