en
Scientific article
English

In‐Situ Spin Labeling of His‐Tagged Proteins: Distance Measurements under In‐Cell Conditions

Published inChemistry, vol. 19, no. 41, p. 13714-13719
Publication date2013-08-26
First online date2013-08-26
Abstract

New spin labeling strategies have immense potential in studying protein structure and dynamics under physiological conditions with electron paramagnetic resonance (EPR) spectroscopy. Here, a new spin‐labeled chemical recognition unit for switchable and concomitantly high affinity binding to His‐tagged proteins was synthesized. In combination with an orthogonal site‐directed spin label, this novel spin probe, Proxyl‐ tris NTA (P‐ tris NTA) allows the extraction of structural constraints within proteins and macromolecular complexes by EPR. By using the multisubunit maltose import system of E. coli : 1) the topology of the substrate‐binding protein, 2) its substrate‐dependent conformational change, and 3) the formation of the membrane multiprotein complex can be extracted. Notably, the same distance information was retrieved both in vitro and in situ allowing for site‐specific spin labeling in cell lysates under in‐cell conditions. This approach will open new avenues towards in‐cell EPR.

eng
Keywords
  • Chelators
  • Electron paramagnetic resonance
  • Membrane proteins
  • Molecular recognition
  • Spin probes
Affiliation Not a UNIGE publication
Citation (ISO format)
BALDAUF, Christoph et al. In‐Situ Spin Labeling of His‐Tagged Proteins: Distance Measurements under In‐Cell Conditions. In: Chemistry, 2013, vol. 19, n° 41, p. 13714–13719. doi: 10.1002/chem.201301921
Main files (1)
Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0947-6539
17views
0downloads

Technical informations

Creation12/21/2023 3:45:18 PM
First validation12/22/2023 10:07:41 AM
Update time12/22/2023 10:07:41 AM
Status update12/22/2023 10:07:41 AM
Last indexation05/06/2024 5:40:12 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack