A Bioresistant Nitroxide Spin Label for In-Cell EPR Spectroscopy: In Vitro and In Oocytes Protein Structural Dynamics Studies.
Angew Chem Int Ed Engl
; 57(5): 1366-1370, 2018 01 26.
Article
in En
| MEDLINE
| ID: mdl-29227566
ABSTRACT
Approaching protein structural dynamics and protein-protein interactions in the cellular environment is a fundamental challenge. Owing to its absolute sensitivity and to its selectivity to paramagnetic species, site-directed spin labeling (SDSL) combined with electron paramagnetic resonance (EPR) has the potential to evolve into an efficient method to follow conformational changes in proteins directly inside cells. Until now, the use of nitroxide-based spin labels for in-cell studies has represented a major hurdle because of their short persistence in the cellular context. The design and synthesis of the first maleimido-proxyl-based spin label (M-TETPO) resistant towards reduction and being efficient to probe protein dynamics by continuous wave and pulsed EPR is presented. In particular, the extended lifetime of M-TETPO enabled the study of structural features of a chaperone in the absence and presence of its binding partner at endogenous concentration directly inside cells.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oocytes
/
Xenopus Proteins
/
Nitrogen Oxides
Limits:
Animals
Language:
En
Journal:
Angew Chem Int Ed Engl
Year:
2018
Document type:
Article
Affiliation country:
Francia