Protein dynamics in living cells studied by in‐cell NMR spectroscopy

Abstract

a b s t r a c tMost proteins function in cells where protein concentrations can reach 400 g/l. However, most quantitative studies of protein properties are performed in idealized, dilute conditions. Recently developed in-cell NMR techniques can provide protein structure and other biophysical properties inside living cells at atomic resolution. Here we review how protein dynamics, including global and internal motions have been characterized by in-cell NMR, and then discuss the remaining challenges and future directions.

Residue level quantification of protein stability in living cells

“…These properties are expected to result in a large number of charge-charge repulsions in cells. Indeed, it has been suggested that these repulsions are what allow GB1 to tumble freely in the cell and yield high quality in-cell 15 N-1 H HSQC spectra (13,14,54,55,67,68). These repulsive interactions enhance the volume excluded by hard interactions in cells, thus favoring the compact native state and resulting in the observed stabilization.…”

Inhibition of protein aggregation and amyloid formation by small molecules

“…While structure determination of small oligomers of Ab/ inhibitor by combining simulations, solution NMR and electron-spray ionization mass-spectrometry [41 ], would represent a real breakthrough, in-cell NMR experiments on amyloid peptides alone and with inhibitors are critical because crowding can change the stability and equilibrium ensembles [42]. It is also important to take into account that the population of brain Ab oligomers changes with aging [43 ], and understand why the A2T and A2V mutations protect from AD [44 ,45].…”

Peptidoglycan Remodeling by the Coordinated Action of Multispecific Enzymes

“…The importance of conformational flexibility for catalytic efficiency has promoted the development of new methodologies such as molecular dynamics and nuclear magnetic resonance that permit investigation of protein mobility at time scales of femtoseconds to seconds. 49,54 …”