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  W-band orientation selective DEER measurements on a Gd3+/nitroxide mixed-labeled protein dimer with a dual mode cavity

Kaminker, I., Tkach, I., Manukovsky, N., Huber, T., Yagi, H., Otting, G., et al. (2013). W-band orientation selective DEER measurements on a Gd3+/nitroxide mixed-labeled protein dimer with a dual mode cavity. Journal of Magnetic Resonance, 227, 66-71. doi:10/1016/j.jmr.2012.11.028.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-8026-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-EF04-6
Genre: Journal Article

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1629151.pdf (Publisher version), 479KB
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 Creators:
Kaminker, I., Author
Tkach, I.1, Author              
Manukovsky, N., Author
Huber, T., Author
Yagi, H., Author
Otting, G., Author
Bennati, M.1, Author              
Goldfarb, D., Author
Affiliations:
1Research Group of Electron Paramagnetic Resonance, MPI for biophysical chemistry, Max Planck Society, ou_578606              

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Free keywords: EPR; DEER; Gd3+ spin labels; Dual mode cavity; High field; Orientation selection; Distance measurements
 Abstract: Double electron–electron resonance (DEER) at W-band (95 GHz) was applied to measure the distance between a pair of nitroxide and Gd3+ chelate spin labels, about 6 nm apart, in a homodimer of the protein ERp29. While high-field DEER measurements on systems with such mixed labels can be highly attractive in terms of sensitivity and the potential to access long distances, a major difficulty arises from the large frequency spacing (about 700 MHz) between the narrow, intense signal of the Gd3+ central transition and the nitroxide signal. This is particularly problematic when using standard single-mode cavities. Here we show that a novel dual-mode cavity that matches this large frequency separation dramatically increases the sensitivity of DEER measurements, allowing evolution times as long as 12 ls in a protein. This opens the possibility of accessing distances of 8 nmand longer. In addition, orientation selection can be resolved and analyzed, thus providing additional structural information. In the case of W-band DEER on a Gd3+– nitroxide pair, only two angles and their distributions have to be determined, which is a much simpler problem to solve than the five angles and their distributions associated with two nitroxide spin labels.

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Language(s): eng - English
 Dates: 2012-12-122013
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10/1016/j.jmr.2012.11.028
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Title: Journal of Magnetic Resonance
Source Genre: Journal
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Pages: - Volume / Issue: 227 Sequence Number: - Start / End Page: 66 - 71 Identifier: ISSN: 0022-2364