5 Years Quantitative Proteomics Course

 

Meet Christina Ludwig (CL), Jeroen Krijgsveld (JK) and Mikhail Savitski (MS) – organisers of the EMBO Practical Course: Quantitative Proteomics: Strategies and Tools to Probe Biology (21 – 26 June 2020). This year marks the course’s 5th anniversary and since 2016 it has grown in popularity and application numbers, reaching 164 applications for 24 seats in 2018. Christina, Jeroen and Mikhail share with us how the course has developed over the years and what their vision is for its future.

 1.  This year marks the 5th anniversary of the Quantitative Proteomics course. Back in 2016, why did you decide to organise it?

JK: The main motivation to initiate the course was because proteomics has become a mature technology that is increasingly being used by biologists to identify proteins, their modifications, interactions etc. However, few biologists have direct access to mass spectrometers, so they use them via collaborators or core facilities. They then get the results in a tabular form, often in a large excel sheet, from which they extract biological interpretation of the experiment. Importantly, we felt that the area between handing in a sample for mass spectrometric analysis and receiving the results was largely a black box. So in the course we aimed to demystify this, and explain the principles and strategies to generate information from raw MS data, and to train them in the use of computational tools to achieve this. Also, we aimed to give insight that proteomics can be done in various ways, so that participants may design their experiments such that they best address the question they are looking to answer. Finally, we aimed to equip participants with some terminology that will help them to communicate with their MS-collaborators, and ask the right questions. Because in many cases proteomics remains a team effort!

2. How has the course developed since?

JK: Proteomics is a very broad field with many mass spectrometric approaches, methods for data analysis and biological applications, making it impossible to cover this in a 1-week course. While in all editions of the course we have maintained a core that explains the main principles in proteomics and covers all of the current state-of-the-art quantitative technologies used in proteomics. Additionally, we have included other elements that varied over the years, to highlight emerging topics or specific application areas, e.g. in structural biology or immunology. This year, we are happy to include a module focusing on statistical analysis of large-scale proteomic data, which is a recurrent issue in almost any proteomics experiment.

3. How do you choose which bioinformatics tools to cover in the course?

JK: There is an increasing number of bioinformatic tools that can analyse the same data using different underlying algorithms. Several of them have matured a lot over the years, making them more robust or have additional functionality. It is not always easy for anyone to know, when looking for an ‘analysis pipeline’, which tool can be best used. It can actually be a bit confusing that the same data can produce different results depending on the tool that is used, while at the same time none will be wrong. So instead of telling which tool is the best, we explain some of the underlying assumptions and the influence one has by choosing certain settings. I think for a researcher it is more important to justify how the data were processed, instead of saying that they used a certain software tool.

4. What could the techniques in this course be used for in the bigger picture?

CL: Proteomics technologies have reached a level of comprehensiveness, throughput and quantitative quality that was inconceivable just a few years back. However, applying proteomics to biological projects still requires lots of knowledge about experimental design, optimal sample preparation, most suitable mass spectrometric technologies and statistical interpretation. If we manage to bring both worlds together and teach biologists about the power, as well as the caveats, of proteomics, I think this will really impact life science in many aspects and truly transform the way how scientific projects are carried out for many scientists all over the world.

JK: I agree. Demonstrating the versatility, and thereby the potential and broad utility of proteomics in different contexts is sometimes an eye-opener for course participants. Actually, it is interesting and useful that participants come from all corners of biology, from paleobiology to clinical biomarker discovery. Having those together in a room for a week and interact, with proteomics as the common interest, is fascinating to see as an organiser. And we explicitly facilitate such interactions in discussion groups – it is an important goal of the course.

5. How do you see this course growing in the future?

CL: I think one special feature of this course, compared to other proteomics courses, is that its rather familial in character due to the small number of 24 participants, and that they come from purposefully different countries and research institutes. This rather small group size is optimal in terms of group dynamics and allows lots of personal exchange between participants and speakers, as well as an optimal support during the practical sessions. Therefore, I hope also in the future the small and familiar atmosphere of this course will remain.

JK: What I also hope, and what we’ll try to achieve, is to remain up-to-date and include novel technologies that are emerging. After 20 years of steep development in mass spectrometry, one would expect that this levels off at some point, but this is not the case at all – it is actually difficult to keep up with what is happening, and with what is possible today that you would not dare to think about yesterday. Therefore, a remaining goal for us is to invite speakers and trainers who work at the forefront of technology, but who can also bridge this to important biological applications. This is what excites us as organisers, and we hope that this will help to make this one of the courses to go to for younger generations of scientists, and get infected too.

6. What motivates you most about your work?

CL: What I really love about heading a proteomics core facility is the huge variety of cool scientific projects you get exposed to, as well as the fact that you work closely with lots of very different scientists coming from completely different scientific disciplines. Every project and every collaboration partner challenges you in terms of diving into a new research area, providing an optimal proteomic workflow and also teaching and educating your collaboration partners in understanding their proteomic data.

MS: The fact that you have the constant possibility to come up and implement creative ideas is incredibly rewarding. Also the fact in research you are constantly generating results that are the first of their kind. There is always an experiment done that has not been done by anyone before and you are the first to see the results. I also love the academic environment the freedom and craziness of it all.

7. Why did you end up in the field of Proteins and Proteomics?

CL: Already during my Chemistry studies all the “biochemistry” lectures and practicals that focused on proteins and life sciences were by far the most interesting subjects for me. During my PhD, which I did in the field of protein engineering at the TU Dortmund, I studied a specific class of proteins, so called inteins, but I hardly applied any mass spectrometry during that time. However, for one specific experiment I used for the first time MALDI-MS to identify the reaction products of a set of purified inteins. My MALDI measurements showed the occurrences of an unexplainable loss of 18 m/z for one of my inteins. First I thought I did a mistake and was very frustrated. But when I repeated and further investigated my samples using also ESI tandem mass spectrometry I could proof the existence of a very interesting cyclic protein-intermediate, which actually helped me explaining the underlying protein splicing mechanism. This turned out being the most interesting result of my whole PhD.

MS: I originally was very focused on pure mathematics. By chance I had an encounter with Roman Zubarev who was a new professor at Uppsala University at the time. His drive, energy and passion for science convinced me to switch fields from mathematics to mass spectrometry and proteomics, which I never regretted.

8. What could you not do without in your life?

CL: Well, as a mother of two beautiful kids the very first thing I could not do without in my life is of course my family :)! And together with my family we love being outdoors, ideally in the Alps, either on (mountain)bikes, rock climbing or hiking. Living without mountains and outdoor activities would be very hard.

MS: First and foremost, my family! Second is physical activity. I love science and I love working a lot, but it takes its toll physically and mentally. My perfect way of recovering and getting the energy back is ideally by rock climbing, running and being out in nature in general.

9. If you would get the chance to meet a famous person – no matter if this person is still alive or not – who would that be?

CL: As a hobby climber I would really like to once meet Alex Honold, who is a world famous free-solo climber who climbed many of the most difficult and exposed climbs in Yosemite National Park without rope. Alex seems in interviews and videos like a really nice and funny guy, but I believe his brain must function very differently than mine when it comes to fear of height, so I would love chat with him about that ;).

MS: I was always interested in mathematics as well as computer science. It would have been fascinating to meet Alan Turing and discuss his vision of how things would develop based on what he knew back then. Incidentally, he was also a really excellent long distance runner with sub 3 hours’ marathon times. It would have been exciting to have a discussion over a run on the countryside :).

10. Which was the best decision in your career so far?

CL: I think the best decision for my career was to perform my Postdoc in the group of Professor Ruedi Abersold at the ETH Zürich, because this has really been the door opener for my career so far. When I finished my PhD it was actually not easy for me to decide for a postdoc in the field of mass spectrometry, because I hardly had any MS experience (I only performed this one MS experiment that I already described above ;)). And starting in a proteomics expert lab as a postdoc who had never really done proteomics before was definitely not easy in the beginning. But I did learn a lot of new things fast and ultimately this allowed me to bring together the two different expertises from my PhD and my Postdoc, which I do believe is a big advantage for any scientific career.

MS: Professionally, I think doing PhD in mass spectrometry was probably the best decision I have made so far. That early in your career, one still knows very little of the world and some luck is definitely required.


Interested in this course? Apply by 22 March!

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Author: Krasimira

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