Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

Abstract

Investigating the structure, modification, interaction, and function of biomolecules in their native cellular environment leads to physiologically relevant knowledge about their mechanisms, which will benefit drug discovery and design. In recent years, nuclear and electron magnetic resonance (NMR) spectroscopy has emerged as a useful tool for elucidating the structure and function of biomacromolecules, including proteins, nucleic acids, and carbohydrates in living cells at atomic resolution. In this review, we summarize the progress and future of in-cell NMR as it is applied to proteins, nucleic acids, and carbohydrates.

DNP‐Supported Solid‐State NMR Spectroscopy of Proteins Inside Mammalian Cells

“…[1][2][3][4] On the other hand, solid-state NMR spectroscopy (ssNMR) has been used to probe proteins and large protein complexes in bacterial cells [5][6][7][8][9] and at the cell membrane periphery of human cells. [1][2][3][4] On the other hand, solid-state NMR spectroscopy (ssNMR) has been used to probe proteins and large protein complexes in bacterial cells [5][6][7][8][9] and at the cell membrane periphery of human cells.…”

DNP‐Supported Solid‐State NMR Spectroscopy of Proteins Inside Mammalian Cells

“…Increasing evidence suggests that the highly complex and dynamic environment of the cell interior and its physiochemical setting imposes critical control on cellular functions, which is hardly reproducible under in vitro conditions.In-cell solution-state NMR spectroscopy can track such structural and dynamic interactions at the atomic level provided that proteins or other molecular units are small and tumble rapidly. [1][2][3][4] On the other hand, solid-state NMR spectroscopy (ssNMR) has been used to probe proteins and large protein complexes in bacterial cells [5][6][7][8][9] and at the cell membrane periphery of human cells. [10] However,extending such studies to investigating proteins and molecular complexes inside human cells poses additional challenges.Firstly,molecule-specific isotope labeling must be achieved to spectroscopically detect the protein of interest in acomplex cellular background.…”

Protein folding and quinary interactions: creating cellular organisation through functional disorder

“…Future experiments need to consider in vivo approaches (NMR , FReI ), where the weak associations between proteins and the differently crowded intracellular environments are preserved. These methods must be complemented with in vitro analysis because the in vivo approaches mask the individual function of crowding molecules .…”