Claudia’s update on her research

This month Claudia shares with us her experice as a RAMP ESR student and an update on her research.

– Could you remind us briefly what your project is about?

I am working on one of the adenosine receptors. This is a membrane receptor translating signals from the outside into the interior of the cell. It belongs to the family of G protein coupled receptors (GPCRs). The different receptors in this family recognize a large variety of signals, ranging from light to neurotransmitters and hormones, but they all have very similar way of translating these signals into the cell.

Interestingly, there are many different signalling outputs for these receptors. One way to modulate these outputs is though the binding of different proteins to the receptor. However, currently very little is known about these interactions. The aim of my project is to determine the structure of the adenosine receptor together with one of these proteins to be able to better characterize the nature of this interaction. A better understanding of these interactions will be crucial to develop more specific drugs which can modulate the receptor to generate only the desired signalling outputs.

– What important milestone have you reached until now?

To be able to study the complex of the adenosine receptor and its binding partners, I first need to produce large amounts of pure protein. This is usually one of the big bottle-necks for membrane protein crystallisation. Very often, the proteins have to be slightly modified to be more stable and tags are added to enable you to fish out your protein of interest out of a cell soup. After designing your protein, you have to introduce the DNA into cells which produce your protein for you. In my case I integrated the DNA of my protein into the genome of a special virus that infects insect cells. The infected cells then produce my protein of interest instead of viral proteins.

I now am able to produce several versions of my protein which I can purify and use to try and form the complex with.

In a parallel approach, I am directly producing both proteins of the complex inside of the same cell. By attaching fluorescent signals to the proteins, I was able to visualise first evidence of complex formation.

– Did the ITN help you in the implementation of your project until now? If yes in what way?

I get a lot of support from our collaborator Dr. Veli-Pekka Jaakola at Novartis. He has a lot of experience in working with this specific receptor and kindly answers all the emails I am bombarding him with. Through this collaboration, I also obtained some additional variants of my receptor and some constructs for a second binding protein.

– Would you recommend other students to apply to a position within a MSCA network such as RAMP? What advice would you give them?

Yes, definitely! I think the MSCA network provides lots of additional opportunities compared to other PhD positions. I really appreciate having a great network of support, consisting of top experts in the field and students which are in the same position as me. I enjoy working on my own project but at the same time having a common goal with all the other fellows. It is exciting to learn about all the different aspects that people with different backgrounds can bring to the same problem.

In particular, the secondments are a great opportunity to get to know different labs and cities and to network with people in the field. However, one must bear in mind that it is also quite stressful to frequently move to different places. It is therefore important to plan the secondments carefully. Valuable time of your PhD must be invested in doing the same safety training etc. at every new institution to comply with the local rules and you have to account some time to get to know the new environment and techniques before you finally can get started with your own work.

– Is there a topic you would like to share/collaborate within the network in relation to your research work?

I think my project, and maybe a few others, differ from most of the projects in the network, as they focus on a particular biological problem rather than on developing novel methods for membrane protein crystallisation. We try to solve or improve the resolution of the structure of one specific membrane protein to gain more knowledge about its function. This at a first sight might have not much to do with “rationalising membrane protein crystallisation”. However, we can provide the “real-life problems“ associated with membrane protein crystallisation and therefore are able to provide lots of data (also valuable negative data) for the generation of novel or improved crystallisation techniques as well as a model system to prove them. I think it’s important to occasionally remind the other members of the network about this.