VEGF-C/VEGFR-2 complex structure took 13 years to solve

Finally our paper was accepted for publication in PNAS (downloadable also from here). I started the project by making the first construct 13 years ago, but it went nowhere for the first 9 years due to insufficient concentration of efforts and a few unlucky choices in the experimental design. I opted for bacterial protein first, but although the refolding worked, it was very inefficient (< 2% of the material was biologically active). In addition, the first and obvious choice to aim at the complex structure of VEGF-C with VEGFR-3 appeared to be a dead end. However, after Veli-Matti took over the project lead, we finally started getting somewhere, although it remained challenging. Repeated attempts with different constructs and crystals had to be done, key changes being the Cys137->Ala mutation and a smart trimming of N- and C-terminus. More...