Pulse EPR distance measurements to study multimers and multimerisation
Abstract
Pulse dipolar electron paramagnetic resonance (PD-EPR) has become a powerful tool for structural biology determining distances on the nanometre scale. Recent advances in hardware, methodology, and data analysis have widened the scope to complex biological systems. PD-EPR can be applied to systems containing lowly populated conformers or displaying large intrinsic flexibility, making them all but intractable for cryo-electron microscopy and crystallography. Membrane protein applications are of particular interest due to the intrinsic difficulties for obtaining high-resolution structures of all relevant conformations. Many drug targets involved in critical cell functions are multimeric channels or transporters. Here, common approaches for introducing spin labels for PD-EPR cause the presence of more than two electron spins per multimeric complex. This requires careful experimental design to overcome detrimental multi-spin effects and to secure sufficient distance resolution in presence of multiple distances. In addition to obtaining mere distances, PD-EPR can also provide information on multimerisation degrees allowing to study binding equilibria and to determine dissociation constants.
Citation
Ackermann , K & Bode , B E 2018 , ' Pulse EPR distance measurements to study multimers and multimerisation ' , Molecular Physics , vol. 116 , no. 12 , pp. 1513-1521 . https://doi.org/10.1080/00268976.2017.1421324
Publication
Molecular Physics
Status
Peer reviewed
ISSN
0026-8976Type
Journal article
Description
This work was supported by funding from the European Union (Marie Curie Actions REA 334496), the Carnegie Trust (70098), the EPSRC (EP/M024660/1) and a Wellcome Trust multi-user equipment grant (099149/Z/12/Z).Collections
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