Nanomolar pulse dipolar EPR spectroscopy in proteins : CuII-CuII and nitroxide-nitroxide cases
Date
2021Grant ID
099149/Z/12/Z
204821/Z/16/Z
RPG-2018-397
BB/R013780/1
Keywords
Metadata
Show full item recordAbstract
The study of ever more complex biomolecular assemblies implicated in human health and disease is facilitated by a suite of complementary biophysical methods. Pulse Dipolar electron paramagnetic resonance Spectroscopy (PDS) is a powerful tool that provides highly precise geometric constraints in frozen solution, however the drive towards PDS at physiologically relevant sub-μM concentrations is limited by the currently achievable concentration sensitivity. Recently, PDS using a combination of nitroxide and CuII based spin labels allowed measuring 500 nM concentration of a model protein. Using commercial instrumentation and spin labels we demonstrate CuII-CuII and nitroxide-nitroxide PDS measurements at protein concentrations below previous examples reaching 500 and 100 nM, respectively. These results demonstrate the general feasibility of sub-μM PDS measurements at short to intermediate distances (~1.5 - 3.5 nm), and are of particular relevance for applications where the achievable concentration is limiting.
Citation
Ackermann , K , Wort , J & Bode , B E 2021 , ' Nanomolar pulse dipolar EPR spectroscopy in proteins : Cu II -Cu II and nitroxide-nitroxide cases ' , Journal of Physical Chemistry B , vol. Articles ASAP . https://doi.org/10.1021/acs.jpcb.1c03666
Publication
Journal of Physical Chemistry B
Status
Peer reviewed
ISSN
1520-6106Type
Journal article
Description
This research was funded, in whole or in part, by the Wellcome Trust (099149/Z/12/Z and 204821/Z/16/Z). A CC BY or equivalent license is applied to the author-accepted manuscript arising from this submission, in accordance with the grant’s open-access conditions. B.E.B. and K.A. acknowledge support by the Leverhulme Trust (RPG-2018-397). J.L.W. was supported by the BBSRC DTP Eastbio. B.E.B. acknowledges equipment funding by BBSRC (BB/R013780/1).Collections
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