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ATP-dependent stabilization and protection of fibroblast growth factor 2

Author:
Rose, Karsten, Gast, Ronald E., Seeger, Anna, Krieglstein, Josef, Klumpp, Susanne
Source:
Journal of biotechnology 2010 v.145 no.1 pp. 54-59
ISSN:
0168-1656
Subject:
adenosine diphosphate, adenosine monophosphate, adenosine triphosphate, alkaline phosphatase, angiogenesis, biotechnology, cell culture, cell proliferation, clinical trials, digestion, elastase, fibroblast growth factor 2, heat, human umbilical vein endothelial cells, in vitro digestion, mutants, neuroprotective effect, nucleosides, plasmin, sodium tripolyphosphate, tissue repair, trypsin
Abstract:
Fibroblast growth factor 2 (FGF2) plays a pivotal role in cell proliferation, angiogenesis and neuroprotection. Several clinical trials using this growth factor in bone regeneration, wound healing and cardioprotection are initiated but the inadequate stability of FGF2 after application is one major problem. Binding of ATP to FGF2 and other growth factors has been demonstrated recently. Here we report that ATP, other nucleoside triphosphates and sodium triphosphate protect FGF2 from trypsin, plasmin and neutrophile elastase digestion in vitro. A molar ratio of 2:1 (ligand/FGF2) is sufficient for these protective effects. ADP shows only little, AMP no stabilizing effect on FGF2 indicating that the number of phosphate residues is important. Protection of FGF2 by ATP can be abolished by the addition of alkaline phosphatase hydrolyzing free and FGF2-bound ATP. The mutant FGF2 (K128A/R129A/K134A/K144A) with strongly reduced ATP-binding capacity revealed no detectable protease resistance after incubation with ATP. Furthermore, a stabilizing effect of ATP on FGF2 could also be demonstrated in cell culture experiments. ATP bound to FGF2 increased FGF2-dependent human umbilical vein endothelial cells proliferation when the growth factor was treated with neutrophile elastase or heat. For the first time these data demonstrate protection of FGF2 by bound ATP, other nucleoside triphosphates or sodium triphosphate from rapid protease digestion. Our data provide new evidence that nucleoside triphosphates are capable of protecting FGF2 and favours such stabilization for various, especially medical applications.
Agid:
282646