Spindler, Philipp E. and Waclawska, Izabela and Endeward, Burichard and Plackrneyer, Joern and Ziegler, Christine and Prisner, Thomas F. (2015) Carr-Purcell Pulsed Electron Double Resonance with Shaped Inversion Pulses. JOURNAL OF PHYSICAL CHEMISTRY LETTERS, 6 (21). pp. 4331-4335. ISSN 1948-7185,
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Pulsed electron paramagnetic resonance (EPR) spectroscopy allows the determination of distances, in the range of 1.5-8 nm, between two spin-labels attached to macromolecules containing protons. Unfortunately, for hydrophobic lipid-bound or detergent-solubilized because of a strongly reduced coherence time of the electron spins. Here membrane proteins, the maximum distance accessible is much lower, we introduce a pulse sequence, based on a Carr-Purcell decoupling scheme on the observer spin, where each pi-pulse is accompanied by a shaped sech/tanh inversion pulse applied to the second spin, to overcome the dipolar interaction, allowing a substantially longer observation time window to be achieved. This increases the upper limit and accuracy of distances that can be determined in membrane protein complexes. We validated the method on a bis-nitroxide model compound and applied this technique to the trimeric betaine transporter BetP. Interprotomer distances as long as 6 nm could be reliably determined, which is impossible with the existing methods.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | NA+/BETAINE SYMPORTER BETP; DISTANCE MEASUREMENTS; SPIN-ECHO; SPECTROSCOPY; TRANSPORT; PROTEINS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie > Prof. Dr. Christine Ziegler |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 06 May 2019 12:25 |
| Last Modified: | 06 May 2019 12:25 |
| URI: | https://pred.uni-regensburg.de/id/eprint/4470 |
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