+検索条件
-Structure paper
タイトル | Use of paramagnetic F NMR to monitor domain movement in a glutamate transporter homolog. |
---|---|
ジャーナル・号・ページ | Nat Chem Biol, Vol. 16, Issue 9, Page 1006-1012, Year 2020 |
掲載日 | 2020年6月8日 |
著者 | Yun Huang / Xiaoyu Wang / Guohua Lv / Asghar M Razavi / Gerard H M Huysmans / Harel Weinstein / Clay Bracken / David Eliezer / Olga Boudker / |
PubMed 要旨 | 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. |
リンク | Nat Chem Biol / PubMed:32514183 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.08 - 3.62 Å |
構造データ | EMDB-20922, PDB-6uwf: EMDB-20923, PDB-6uwl: |
化合物 | ChemComp-ASP: ChemComp-6OU: |
由来 |
|
キーワード | TRANSPORT PROTEIN / aspartate transporter |