PMID- 24038571
OWN - NLM
STAT- MEDLINE
DCOM- 20140530
LR  - 20131004
IS  - 1521-3765 (Electronic)
IS  - 0947-6539 (Linking)
VI  - 19
IP  - 41
DP  - 2013 Oct 4
TI  - In-situ spin labeling of his-tagged proteins: distance measurements under in-cell 
      conditions.
PG  - 13714-9
LID - 10.1002/chem.201301921 [doi]
AB  - 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-trisNTA (P-trisNTA) 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.
CI  - Copyright (c) 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
FAU - Baldauf, Christoph
AU  - Baldauf C
AD  - Institute of Biochemistry, Biocenter, Cluster of Excellence-Macromolecular 
      Complexes (CEF-MC), Goethe-University Frankfurt, Max-von-Laue Str. 9, 60438 
      Frankfurt a.M. (Germany).
FAU - Schulze, Katrin
AU  - Schulze K
FAU - Lueders, Petra
AU  - Lueders P
FAU - Bordignon, Enrica
AU  - Bordignon E
FAU - Tampe, Robert
AU  - Tampe R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130826
PL  - Germany
TA  - Chemistry
JT  - Chemistry (Weinheim an der Bergstrasse, Germany)
JID - 9513783
RN  - 0 (Proteins)
RN  - 0 (Spin Labels)
RN  - 69-79-4 (Maltose)
SB  - IM
MH  - Electron Spin Resonance Spectroscopy/methods
MH  - Escherichia coli/*chemistry/metabolism
MH  - Maltose/*chemistry/metabolism
MH  - Protein Binding
MH  - Proteins/*chemistry/metabolism
MH  - Spin Labels
OTO - NOTNLM
OT  - chelators
OT  - electron paramagnetic resonance
OT  - membrane proteins
OT  - molecular recognition
OT  - spin probes
EDAT- 2013/09/17 06:00
MHDA- 2014/05/31 06:00
CRDT- 2013/09/17 06:00
PHST- 2013/05/18 00:00 [received]
PHST- 2013/09/17 06:00 [entrez]
PHST- 2013/09/17 06:00 [pubmed]
PHST- 2014/05/31 06:00 [medline]
AID - 10.1002/chem.201301921 [doi]
PST - ppublish
SO  - Chemistry. 2013 Oct 4;19(41):13714-9. doi: 10.1002/chem.201301921. Epub 2013 Aug 
      26.