A gadolinium spin label with both a narrow central transition and short tether for use in double electron electron resonance distance measurements
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The design, synthesis, and application of a nine-coordinate gadolinium(III)-containing spin label, [Gd.sTPATCN]-SL, for use in nanometer-distance measurement experiments by EPR spectroscopy is presented. The spin label links to cysteines via a short thioether tether and has a narrow central transition indicative of small zero-field splitting (ZFS). A protein homodimer, TRIM25cc, was selectively labeled with [Gd.sTPATCN]-SL (70%) and a nitroxide (30%) under mild conditions and measured using the double electron electron resonance (DEER) technique with both commercial Q-band and home-built W-band spectrometers. The label shows great promise for increasing the sensitivity of DEER measurements through both its favorable relaxation parameters and the large DEER modulation depth at both Q- and W-band for the inter-Gd(III) DEER measurement which, at 9%, is the largest recorded under these conditions.
Shah , A , Roux , A , Starck , M , Mosely , J A , Stevens , M , Norman , D G , Hunter , R I , El Mkami , H , Smith , G , Parker , D & Lovett , J E 2019 , ' A gadolinium spin label with both a narrow central transition and short tether for use in double electron electron resonance distance measurements ' , Inorganic Chemistry , vol. Article ASAP . https://doi.org/10.1021/acs.inorgchem.8b02892
Copyright © 2019 American Chemical Society. 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 may differ slightly from the final published version. The final published version of this work is available at 10.1021/acs.inorgchem.8b02892
DescriptionFunding from ERC (266804), EPSRC (EP/R013705/1, EP/M508214/1), Carnegie Trust (RIG007510), Wellcome Trust for a Multi-User Equipment Grant (099149/Z/12/Z), and the Royal Society for a University Research Fellowship to JEL.
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