TY  - JOUR
A1  - Pintér, György
A1  - Schwalbe, Harald
T1  - Refolding of cold-denatured barstar induced by radio-frequency heating: a new method to study protein folding by real-time NMR spectroscopy
T2  - Angewandte Chemie
N2  - The C40A/C82A double mutant of barstar has been shown to undergo cold denaturation above the water freezing point. By rapidly applying radio-frequency power to lossy aqueous samples, refolding of barstar from its cold-denatured state can be followed by real-time NMR spectroscopy. Since temperature-induced unfolding and refolding is reversible for this double mutant, multiple cycling can be utilized to obtain 2D real-time NMR data. Barstar contains two proline residues that adopt a mix of cis and trans conformations in the low-temperature-unfolded state, which can potentially induce multiple folding pathways. The high time resolution real-time 2D-NMR measurements reported here show evidence for multiple folding pathways related to proline isomerization, and stable intermediates are populated. By application of advanced heating cycles and state-correlated spectroscopy, an alternative folding pathway circumventing the rate-limiting cis-trans isomerization could be observed. The kinetic data revealed intermediates on both, the slow and the fast folding pathway.
KW  - barstar
KW  - proline isomerization
KW  - NMR spectroscopy
KW  - protein folding
KW  - temperature jump
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/63844
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-638447
SN  - 1521-3773
N1  - This work was supported by Deutsche Forschungsgemeinschaft (GRK 1986 “CLiC”) and European Union Horizon 2020 Program (Marie Sklodowska-Curie Grant 642773). Open access funding enabled and organized by Projekt DEAL.
VL  - 59
IS  - 49
SP  - 22086
EP  - 22091
PB  - Wiley-VCH
CY  - Weinheim
ER  -