1. Visualizing Angiogenesis A CREST Initiative Elise A. Pellmann – Marquette University Daniel Sem, Ph.D. – Concordia University Wisconsin David Goodsell, Ph.D. – Scripps Research Institute

2. 2. Movie 1. Cellular landscape

3. 2 x 10 7 magnification Show: MAPK and receptor tyrosine kinase signaling VEGF, plurality of biochemical cascades angiogenesis cell permeability

4. Knowledge needed for landscape pathway: which proteins? size, domain arrangement/shape, assembly, modifications, interactions/function

5. Pathway: KEGG

6. Pathway

7. Proteins (i)

8. Proteins (ii)

9. Sketches

10. Final version

11. 2. Movie PyMOL, 3D Show: drug binding to DUSP-5

12. DUSP-5 ERK-2 phosphatase: P-T183, P-Y185 C-terminal phosphatase domain N-terminal ERK-binding domain linker region Ser to Pro mutation (hemangiomas)

13. DUSP-5 model C N catalytic domain ERK-binding domain + + – – C: PDB 2G6Z N: Homology model, NMR Linker: hand- drawn Domain orientation: good guess?

14. DUSP-5 model C N catalytic domain ERK-binding domain + + – – 1.ERK binding 2.catalytic domain “clamps” down 3.phosphates removed from ERK Drug impedes catalytic action, may also prevent ERK binding if DUSP-5 clamps down on drug

15. Daniel Sem, Terrence Neumann, Prachi Pradeep David Goodsell Margaret Franzen, MSOE CREST grant Movie viewing Thank you