Protein Structural Dynamics in Living Cells - Project Summary/Abstract
Our current understanding of in vitro protein folding is due to decades of experimental and computational
research that provided high-resolution characterization of protein structure, identification of folding principles,
and development of folding algorithms. However, proteins often participate in new and unexpected functional
and pathological behaviors in vivo. Because protein processes involve a large network of interactions that
strongly depend on the environment, understanding how proteins work inside cells requires knowledge of protein
structure, stability, and dynamics in vivo. While evidence that the cellular environment perturbs protein behaviors
emerged over half a century ago, we still have limited fundamental information about the effects of these
cooperative cellular interactions on protein properties. The gap in knowledge is largely attributable to the weak
transient nature of interactions in the cellular milieu and challenges associated with studying protein functions in
living cells. This limitation is concerning because proteins in cells and organisms are continuously interacting
with other biomolecules, which may disrupt the ability of a protein to fold and assemble properly and results in
loss of function and eventually disease. To address these gaps, our research group leverages groundbreaking
in vivo spectro-microscopy methods, in combination with functional biochemical assays, in vitro biophysical
spectroscopy, and numerical analysis solutions to characterize protein structural dynamics in living cells and
tissues. This platform will transform our ability to examine unexplored layers of protein complexity and regulation
in cells and tissues, specifically: (1) Do classic in vitro protein principles translate to cells? How accurate are the
in-cell predictions of folding theory and molecular dynamics simulations? (2) Can we develop methods to
visualize the spatial distribution of metabolism and associated metabolic protein structural dynamics in living
cells? (3) How does thermal adaptation and acclimation by organisms change the stability, folding, and
aggregation of proteins in differentiated tissues? Overall, this work will lead to a greater understanding of how
remodeling of the cell interior during development, environmental stress, and disease contributes to protein
homeostasis. Unraveling these interactions will improve our molecular-level understanding of essential
processes in human health and disease.