Stage Master

One of the long-term interests of the Wilmanns group resides in the activity regulation of calcium/calmodulin (Ca2+/CaM)-dependent protein kinases. In the last five years, we have contributed to unravel the structure of the pro-apoptotic death-associated protein kinase 1 (DAPK1), in the presence of the regulatory protein Ca2+/CaM (de Diego et al., 2010). This structure provided insight into how Ca2+/CaM binding and other structural changes lead to DAPK activation (Temmerman et al., 2013&2014). More recently, a dimeric DAPK2 crystal structure has also revealed the importance of DAPK2 dimerisation for Ca2+/CaM binding regulation (Simon et al., 2016).
Based on the structures of DAPK1 and DAPK2 in complex with Ca2+/CaM, we aim to mechanistically understand the conformational implications of these post-translational modifications in the release of auto-inhibition. We also want to solve the structure of other kinases in complex with Ca2+/CaM or to trap the known complexes in alternative conformations.
The master student will be supervised by Anne-Sophie Huart, a postdoc in the Wilmanns group since nearly 3 years (https://www.researchgate.net/profile/Anne-Sophie_Huart). English is the used language at EMBL and Hamburg is an international city. The candidate, who would already have some related experience, will mainly focus on cloning (restriction-based and SLiCE cloning, mutagenesis), expression (E.coli cultures, eventually use of insect cell pellets) and purification of different DAPK variants (affinity-based techniques, size exclusion columns, on the bench and using AEKTAs). Using successfully purified proteins, the student would assist in their biophysical characterisations (thermofluor analysis, circular dichroism, dynamic light scattering) and binding assays with Ca2+/CaM by isothermal titration calorimetry and fluorescence anisotropy.