Cross-talk between sphingolipid and phosphoprotein signaling pathways

SPHINGONET-ESR3

Project description
This project will investigate the unexplored convergence of lipid and phosphorylation mediated signaling by both targeted and global phosphoproteomics. Initial goal is to meticulously identify phosphorylation events occurring on several crucial sphingolipid signaling hubs in yeast. Quantitative phosphoproteomics will be used in combination with TAP-tagged signaling hub purification under different sphingolipid conditions. Knock out strains will subsequently be screened to identify responsible kinases and phosphatases. In a mouse model of NPC disease, global phosphoproteomics screens will be performed. First in a cell model where U18666A treated cells will be quantitatively compared with untreated cells. In addition, tissues of NPC1 knock-out mice will be quantitatively compared to wild-type mice. Specific findings from these models will be followed up in patient material of NPC disease.

Further reading
Boersema et al. 2009, Nat Protoc. 4: 484-494

Host institute
Utrecht University, Department of Chemistry
Biomolecular Mass Spectrometry & Proteomics
Supervisors: Dr. Arjen Scholten / Prof. Albert Heck
E-mail: a.scholten@uu.nl
Phone: +31(0)302536793
Web: bioms.chem.uu.nl/

The Biomolecular Mass Spectrometry and Proteomics Group led by prof. Albert Heck houses over 20 mass spectrometers for the analysis of peptides, proteins and protein complexes. The group forms the core of the Netherlands Proteomics Center and is coordinator of the FP7 project ‘PRIME-XS’, aimed at providing access to state-of-the-art proteomics facilities to European Life Science researchers. Research of the group focuses on the development and implementation of innovative mass spectrometric methods for a more efficient and detailed characterization of proteins in relation to their biological function. The emphasis is on the structural characterization of proteins, their post-translational modifications, as well as on protein complexes and protein interaction networks. The lab is fully equipped for the comparative analysis of complete (phospho)proteomes as well as for the characterization of protein interactomes of signalling molecules, lipids or other biomolecules using chemical proteomics strategies. In addition, signaling pathways can be studied by specifically analyzing (de-)phosphorylation events following activation, using in-house developed unique enrichment technologies to isolate phosphopeptides from complex mixtures in combination with dedicated fragmentation methods like ETD and HCD, which are particularly suitable for the characterization of phosphopeptides. Current methods even allow us to do stimulation-dependent quantitative phosphorylation analysis directly in tissue. From a structural point of view, the lab has unique experience with the analysis of intact protein complexes by native mass spectrometry and ion mobility as well the determination of the stoichiometry and shape of the protein complexes involved. As such, the lab forms one of the nodes of the Associate Center for Mass Spectrometry of Native Proteins and Complexes of the FP7-funded INSTRUCT consortium, which is part of the ESFRI roadmap for Research Infrastructures.

Requirements
•    We are looking for an ambitious and interactive individual with MSc degree in analytical (bio)chemistry, molecular cell biology or chemical biology. A solid background in yeast biology is an advantage.
•    The candidate will be part of a multidisciplinary team of chemists, geneticists, biophysicists, biochemists and cancer biologists. Good communication skills are essential.
•    Proactive participation in SPHINGONET’s research & training program is expected through presentations and critical thinking at internal scientific forums, requiring excellent English presentation and writing skills. The ability to work in a team is a must.
•    Candidates must fulfill the eligibility criteria for Marie Curie Initial Training Networks (FP7).
•    Candidates should submit the complete application package.
•    Suitable candidates will be invited for an interview by the supervisor of the project.

Appointment
•    PhD student for 3 years. Extension of the contract to 4 years is possible.
•    Full-time (42 hours/week).