Research Lab Profile: Anand Group
Ganesh Anand, associate professor of chemistry and biochemistry & molecular biology, talks about his lab group and the impact Covid-19 had on their research of protein dynamics.
Kathryn: You joined the Penn State Chemistry faculty in 2020, which was the beginning of the global Covid-19 pandemic. How were you able to adapt to a new position with new colleagues, create the foundations for a successful research lab and ground yourself in an unfamiliar place all amid the uncertainty from the pandemic? What resources in the department were most helpful to you in this transition?
Ganesh: Moving to Penn State at the end of 2020 was certainly disorienting due to the pandemic. It was not a typical busy campus and there were few people to interface with in person. The logistics of the move were harder and instrument deliveries were much slower than normal. But I do want to thank my Penn State Chemistry colleagues who made the transition a lot easier. I specifically wish to acknowledge Phil Bevilacqua, Paul Cremer, Sarah Lonsinger, and Pam Wagner amongst the many colleagues who never hesitated to meet in person whenever I needed to, especially during a time of social distancing with rapidly evolving rules for in-person meetings! In addition, my mentors in the department were always available for ZOOM calls. People will always be the best resources in any workplace and my chemistry colleagues were the best resources for my career transition at an unusual period in the world.
Kathryn: What projects are you and your group members currently working on?
Ganesh: My group is working on determining the dynamics at seconds and slower time scales of large macromolecular complexes such as viruses and signaling enzymes. Our target viruses are Coxsackie, Dengue, SARS CoV-2, and Turnip Crinkle virus. We seek to probe fluctuations in virus conformations, termed “breathing”. Virus breathing is an essential feature that assists in their ability to seek host cells for binding and entry during infection. A few members are also examining the phenomenon of ligand channeling in large multi-subunit signaling enzyme complexes.
Kathryn: Your group recently published a paper in the journal eLife on how mutations made SARS-CoV-2 spike proteins more rigid, potentially improving the virus’ fitness. How did the Covid-19 pandemic and continuously emerging SARS-CoV-2 variants impact your initial interest and research on protein dynamics?
Ganesh: Just after my first mass spectrometry instrument was installed in December 2020, I got an opportunity to compare the poses of mutant spike proteins to the original strain in December 2019. At that time there were only a few mutations in SARS CoV-2. Between then and 2022, the world encountered waves of infection beginning with the alpha variant and most recent omicron variant. Each of these variants showed clusters of mutations throughout the spike protein. It was clear that the spike protein was rapidly evolving, and we decided to chronicle how these mutations were altering the pose of spike. What we found was astonishing; a spike protein bundle of three monomers in every variant was more tightly wound up than the spike from a predecessor variant! This indicated that mutations on the surface of spikes were having deeper long-range effects on the protein. This gave us enormous insights into the selection pressure on the spike protein and explained why new variants were able to cause breakthrough infections in individuals treated with vaccines generated against spikes from the original SARS CoV-2 strain.
Kathryn: Did you or your group members encounter any specific difficulties working with the covid virus? What was most surprising about the results you found?
Ganesh: Due to safety considerations, it was not possible to work with the actual SARS CoV-2 virus. Instead, we worked with a recombinant spike protein expressed in mammalian cells. This protein is completely safe to handle for our research, as it is only a non-infectious component of SARS CoV-2. The most surprising find has been why SARS CoV-2 has thrown up so many new variants that are driving a tightening of the spike protein. None of the previous recent coronavirus outbreaks (SARS CoV-1 and MERS) showed variant emergence, which have greatly exacerbated the human costs of this pandemic. We are currently seeking to expand our analysis of the spike from MERS and SARS CoV-1 to identify features that are unique to the spike from SARS CoV-2.
Kathryn: Can you describe some of the interesting research techniques and equipment your group uses? Why are these state-of-the-art tools important and critical to your research?
Ganesh: We have been applying mass spectrometry to probe the dynamics of viruses and signaling complexes. Mass spectrometry has been largely applied to identify and quantitate molecules. Recently, it has been applied to determine structures of molecules in solution and uncover dynamics of molecules, their interactions, and long-range conformational changes. We are fortunate in housing 3 state-of-the-art mass spectrometry instrumentation: an Electrospray ionization quadrupole-Time of flight (ESI-QTof) Synapt X-S (Waters) with Leap robot, Orbitrap Eclipse, and Orbitrap Ultra High Mass Range (UHMR) (Thermo) instruments. Thus far, only static snapshots of viruses and large molecular complexes are available from cryo-electron microscopy and crystallography – a photographic perspective. Structural mass spectrometry is critical to observe how these molecules move and morph in different environments. This is a movie perspective that is critical to determining chemistry in motion and molecular function.
Kathryn: You work with a large and diverse group of motivated graduate researchers. What has been one of your proudest moments as a PI?
Ganesh: My proudest moments as a PI have been to watch the empowerment of graduate researchers as they grow from tentative new members to confident and independent researchers. It is also great to see my students present at international conferences and being lauded for their research, and in some instances, being presented with postdoctoral employment opportunities. Diversity enriches all of us and I am very fortunate to mentor a rich and diverse group of researchers.
Kathryn: What goals do you have for the future of your research group?
Ganesh: My research group seeks to develop a program for determining virus breathing and assembly; the disassembly transitions of viruses through a combination of mass spectrometry, cryo-electron microscopy, and computational approaches. This will uncover the molecular pathway driving the entire lifecycle of viruses. A disassembling virus represents a vulnerable stage in its lifecycle, and we hope our research will guide the design of antibodies that specifically target disassembly hotspots on a virus surface.
