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-Structure paper
| Title | Use of paramagnetic F NMR to monitor domain movement in a glutamate transporter homolog. |
|---|---|
| Journal, issue, pages | Nat Chem Biol, Vol. 16, Issue 9, Page 1006-1012, Year 2020 |
| Publish date | Jun 8, 2020 |
 Authors | Yun Huang / Xiaoyu Wang / Guohua Lv / Asghar M Razavi / Gerard H M Huysmans / Harel Weinstein / Clay Bracken / David Eliezer / Olga Boudker /   |
| PubMed Abstract | In proteins where conformational changes are functionally important, the number of accessible states and their dynamics are often difficult to establish. Here we describe a novel F-NMR spectroscopy ...In proteins where conformational changes are functionally important, the number of accessible states and their dynamics are often difficult to establish. Here we describe a novel F-NMR spectroscopy approach to probe dynamics of large membrane proteins. We labeled a glutamate transporter homolog with a F probe via cysteine chemistry and with a Ni ion via chelation by a di-histidine motif. We used distance-dependent enhancement of the longitudinal relaxation of F nuclei by the paramagnetic metal to assign the observed resonances. We identified one inward- and two outward-facing states of the transporter, in which the substrate-binding site is near the extracellular and intracellular solutions, respectively. We then resolved the structure of the unanticipated second outward-facing state by cryo-EM. Finally, we showed that the rates of the conformational exchange are accessible from measurements of the metal-enhanced longitudinal relaxation of F nuclei. |
 External links | Nat Chem Biol / PubMed:32514183 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.08 - 3.62 Å |
| Structure data | EMDB-20922, PDB-6uwf: EMDB-20923, PDB-6uwl: |
| Chemicals |  ChemComp-ASP:  ChemComp-6OU: |
| Source |
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 Keywords |  TRANSPORT PROTEIN / aspartate transporter |
