Does the cellular environment significantly alter protein structure?

Determine whether and to what extent the crowded, non-ideal cellular environment significantly alters the three-dimensional structures of proteins relative to structures characterized under non-physiological in vitro conditions.

Background

Most experimentally determined protein structures have been solved under non-physiological, in vitro conditions (e.g., crystallography, solution NMR, cryo-EM), whereas proteins function inside crowded cells with substantial macromolecular content and complex interactions. The paper emphasizes that despite extensive structural databases for in vitro conditions, only a few high-resolution in vivo structures exist, leaving uncertainty about how cellular context influences protein conformation.

Förster resonance energy transfer (FRET) and in-cell NMR have begun to probe protein structure in living cells, but available data remain sparse. The authors motivate FRET-assisted molecular dynamics as a path toward atomic-scale modeling in vivo, underscoring the foundational question of whether the cellular environment materially impacts protein structure.