Gast, Ronald E. ; König, Simone; Rose, Karsten ; Ferenz, Katja ; Ferenz, Katja Bettina ; Krieglstein, Josef: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cel [...]
Inhalt
- Methods
- Materials
- Production and purification of recombinant human VEGF-A165
- Labeling of VEGF-A165 with [γ-32P]ATP and [α-32P]ATP
- Plasmin digestion of VEGF-A165 labeled with ATP
- Circular dichroism (CD) spectroscopy
- MALDI-TOF MS
- Cell proliferation assay
- Luminometric measurement of extracellular ATP
- Statistical analysis
- Results
- Binding of ATP to VEGF-A165
- MALDI-TOF MS of the VEGF-A165-ATP complex
- ATP induces a conformational change of VEGF-A165
- Binding of ATP does not protect VEGF-A165 from plasmin cleavage
- Mitogenic activity of VEGF-A165 on HUVECs requires ATP
- Discussion
- Conclusions
- Acknowledgements
- Author details
- Authors' contributions
- References