Pulse EPR distance measurements to study multimers and multimerisation
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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.
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
© 2018 Informa UK Limited, trading as Taylor & Francis Group. This work has been made available online in accordance with the publisher’s policies. This is the author created accepted version manuscript following peer review and as such may differ slightly from the final published version. The final published version of this work is available at https://doi.org/10.1080/00268976.2017.1421324
DescriptionThis 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).
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