6UWF

GltPh in complex with L-aspartate and sodium ions in outward-facing state

Summary for 6UWF

• Entry DOI: 10.2210/pdb6uwf/pdb
• EMDB information: 20922
• Descriptor: Glutamate transporter homolog, ASPARTIC ACID, [(2~{R})-1-[2-azanylethoxy(oxidanyl)phosphoryl]oxy-3-hexadecanoyloxy-propan-2-yl] (~{Z})-octadec-9-enoate (3 entities in total)
• Functional Keywords: aspartate transporter, transport protein
• Biological source: Pyrococcus horikoshii
• Total number of polymer chains: 1
• Total formula weight: 46239.05

Authors

Wang, X.,Boudker, O. (deposition date: 2019-11-05, release date: 2020-06-03, Last modification date: 2024-12-25)

Primary citation

Huang, Y.,Wang, X.,Lv, G.,Razavi, A.M.,Huysmans, G.H.M.,Weinstein, H.,Bracken, C.,Eliezer, D.,Boudker, O.
Use of paramagnetic 19 F NMR to monitor domain movement in a glutamate transporter homolog.
Nat.Chem.Biol., 16:1006-1012, 2020

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 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.

PubMed: 32514183
DOI: 10.1038/s41589-020-0561-6

Experimental method

ELECTRON MICROSCOPY (3.08 Å)