Improved Sensitivity for Long-Distance Measurements in Biomolecules: Five-Pulse Double Electron–Electron Resonance
Open Access
- 21 December 2012
- journal article
- research article
- Published by American Chemical Society (ACS) in The Journal of Physical Chemistry Letters
- Vol. 4 (1), 170-175
- https://doi.org/10.1021/jz301788n
Abstract
We describe significantly improved long-distance measurements in biomolecules by use of the new multipulse double electron–electron spin resonance (DEER) illustrated with the example of a five-pulse DEER sequence. In this sequence, an extra pulse at the pump frequency is used compared with standard four-pulse DEER. The position of the extra pulse is fixed relative to the three pulses of the detection sequence. This significantly reduces the effect of nuclear spin-diffusion on the electron-spin phase relaxation, thereby enabling longer dipolar evolution times that are required to measure longer distances. Using spin-labeled T4 lysozyme at a concentration less than 50 μM, as an example, we show that the evolution time increases by a factor of 1.8 in protonated solution and 1.4 in deuterated solution to 8 and 12 μs, respectively, with the potential to increase them further. This enables a significant increase in the measurable distances, improved distance resolution, or both.Keywords
Funding Information
- National Institute of General Medical Sciences
- National Institute of Biomedical Imaging and Bioengineering
- National Institutes of Health
- National Center for Research Resources
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