FPOP Star

Did you always want to be a scientist?

No. In middle school, I thought I would be a lawyer. And in high school, I thought I’d go into medicine. At University, I majored in biochemistry with the ultimate intention of applying to medical school. But I had to do some research as part of that major, and found it much more interesting than medicine! So that’s how I ended up doing a PhD in science – there was no real “aha” moment, I just did what interested me.

What attracted you to the area of research that you work in now?

It’s been a long journey. I took the first steps at college, where I fell in love with proteins. Protein structure, protein folding – it all fascinated me, so when I began looking for graduate positions, I focused on protein structure laboratories. I ended up working for Jenny Yang at Georgia State University, where we employed a variety of techniques from biochemistry, analytical chemistry and biophysics; for example, fluorescence, circular dichroism, and NMR. We were working on small model proteins, but my interests increasingly gravitated towards large protein complexes. To pursue that interest, I joined the lab of Peter Prevelige (Department of Microbiology, University of Alabama, Birmingham), who was working on virus capsids using advanced techniques, including hydrogen-deuterium exchange MS. And that was a revelation for me; I didn’t know you could do structural studies with MS, until then. After working with Peter, I joined Michael Gross’s laboratory – and that opened up a whole new world! Not only had Mike been using MS for decades – which allowed me to benefit from his wealth of experience – but he had also begun using a new technique called fast photochemical oxidation of proteins (FPOP). It seemed to me that it would address my interests better than conventional MS could, so I started using FPOP for the analysis of large proteins; for example, we were the first to report epitope mapping using FPOP. Then I began thinking about using FPOP for the analysis of very complex systems – and that is how I got into the analytical optimization and method development work that I pursue today. As I said before, I didn’t plan to end up where I am – I just followed what I found interesting!

Do you think your career path has given you a different perspective on analytical method development?

Definitely. Being able to draw on both biochemistry and analytical chemistry helps me to align method development with the unique challenges of biology. For example, cellular processes occur on different time scales, some of which – like signaling cascades and protein folding – are very fast. Conventional flow systems are too slow to capture these processes, so I started developing more high-throughput systems in well-plate formats. Having a foot in both camps has been very helpful – I can cover a range of technologies without necessarily having to seek collaborators.

Your work analyzing protein complexes in cells has been described as “groundbreaking” – is your unusual perspective necessary for this type of endeavor?

Well, it doesn’t seem unusual to me – I just like to try things that nobody has done. It can be a little scary at times, but for the most part it’s fun – pushing boundaries is exciting. I always tell graduate students who want to join my lab that they won’t find any protocols to follow – they have to make the protocols. Embracing this philosophy requires a degree of fearlessness – there’s no real safety net when you’re trying things for the first time. The graduate students who succeed in my laboratory enjoy the challenge of innovation; they like trying something new, and they do it fearlessly. On the other side of the coin, I’ve had some people leave the lab because they cannot operate that way, and I understand that.

Do you anticipate a greater role for automation in the coming years?

In my research, definitely. In fact, we’re about to publish a paper on automation of the FPOP method. In general, automated systems speed up our analysis, which help us study cellular processes. Also, we’ve learnt from Jenny van Eyk’s approach to automating sample processing by means of a Beckman liquid handling device; we are now trying to couple this with the Thermo Scientific 96-well plate system. The idea is to automate sample processing from cell lysis all the way through to sample digestion for MS, and we are working with Thermo Scientific to achieve this. Automated sample processing should improve reproducibility of MS outputs; we want our samples to be as good as possible so that we can get the most out of our MS methods. In brief, anybody engaged in proteomic analysis of biological samples will benefit from process automation, as it improves throughput and, by removing manual errors, offers better reproducibility. In short, I think automation will be hugely important in the continued development of the MS field.

Who has inspired you in the professional domain?

I could give you a long list of inspirational people! But it would have to include Carol Robinson, Vicki Wysocki, and Jenny Brodbelt, who together paved the way for women in the analytical sciences. It’s always been a male-dominated field but that didn’t stop them: Vicky Wysocki developed surface-induced dissociation (SID), Jenny Broadbelt added ultraviolet photodissociation (UVPD) lasers to MS systems, and Carol Robinson took unprecedented steps, such as putting an entire membrane organelle into the mass spectrometer.

What kind of interests do you have outside work?

I really love architecture; (I once thought about majoring in architecture, but I can’t draw!) My interest started in the New York suburbs where I grew up; we have some beautiful Gothic style buildings, like St. Patrick’s Cathedral and the church at Syracuse University. Those edifices captured my attention by virtue of their craftsmanship; there are details in the woodwork, for example, that you just don’t see in modern buildings. My parents’ house had some of the same feeling – it was built in the 1840s, with wonderful woodwork and stained-glass windows. So perhaps my interest in architecture stemmed from the kind of house I grew up in.

Do you have a particular mission for the next five or ten years?

Two things. First, I want FPOP to become accepted as a standard structural biology tool. Second, I want to facilitate training of underrepresented minorities in science. Those are my two key ambitions for the future.