title: Extending the distance range accessed with continuous wave EPR with Gd3+ spin probes at high magnetic fields.
abstract: Interspin distances between 0.8 nm and 2.0 nm can be measured through the dipolar broadening of the continuous wave (cw) EPR spectrum of nitroxide spin labels at X-band (9.4 GHz, 0.35 T). We introduce Gd(3+) as a promising alternative spin label for distance measurements by cw EPR above 7 Tesla, where the |-1/2〉 to |1/2〉 transition narrows below 1 mT and becomes extremely sensitive to dipolar broadening. To estimate the distance limits of cw EPR with Gd(3+), we have measured spectra of frozen solutions of GdCl3 at 8.6 T (240 GHz) and 10 K at concentrations ranging from 50 mM to 0.1 mM, covering a range of average interspin distances. These experiments show substantial dipolar broadening at distances where line broadening cannot be observed with nitroxides at X-band. This data, and its agreement with calculated dipolar-broadened lineshapes, show Gd(3+) to be sensitive to distances as long as ∼3.8 nm. Further, the linewidth of a bis-Gd(3+) complex with a flexible ∼1.6 nm bridge is strongly broadened as compared to the mono-Gd(3+) complex, demonstrating the potential for application to pairwise distances. Gd-DOTA-based chelates that can be functionalized to protein surfaces display linewidths narrower than aqueous GdCl3, implying they should be even more sensitive to dipolar broadening. Therefore, we suggest that the combination of tailored Gd(3+) labels and high magnetic fields can extend the longest interspin distances measurable by cw EPR from 2.0 nm to 3.8 nm. cw EPR data at 260 K demonstrate that the line broadening remains clear out to similar average interspin distances, offering Gd(3+) probes as promising distance rulers at temperatures higher than possible with conventional pulsed EPR distance measurements.
authors.affiliation: Department of Physics, University of California, Santa Barbara, Santa Barbara, California 93106, USA.
authors.forename: Devin T
authors.initials: DT
authors.lastname: Edwards
authors.forename: Zhidong
authors.initials: Z
authors.lastname: Ma
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Meade
authors.forename: Daniella
authors.initials: D
authors.lastname: Goldfarb
authors.forename: Songi
authors.initials: S
authors.lastname: Han
authors.forename: Mark S
authors.initials: MS
authors.lastname: Sherwin
mesh_terms: D002614:Chelating Agents / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D005682:Gadolinium / Q000737:chemistry*; D060526:Magnetic Fields; D009942:Organometallic Compounds / Q000737:chemistry*
chemical_list: D002614:Chelating Agents; D009942:Organometallic Compounds; D005682:Gadolinium