6V8G

GltPh mutant - Y204L A345V V366A

Summary for 6V8G

• Entry DOI: 10.2210/pdb6v8g/pdb
• Descriptor: Glutamate transporter homolog, SODIUM ION, ASPARTIC ACID (3 entities in total)
• Functional Keywords: aspartate transporter, substrate bound, fast mutant, membrane protein
• Biological source: Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
• Total number of polymer chains: 3
• Total formula weight: 134382.01

Authors

Boudker, O.,Huysmans, G.H.M. (deposition date: 2019-12-11, release date: 2020-11-18, Last modification date: 2023-10-11)

Primary citation

Huysmans, G.H.M.,Ciftci, D.,Wang, X.,Blanchard, S.C.,Boudker, O.
The high-energy transition state of the glutamate transporter homologue GltPh.
Embo J., 40:e105415-e105415, 2021

PubMed Abstract: Membrane transporters mediate cellular uptake of nutrients, signaling molecules, and drugs. Their overall mechanisms are often well understood, but the structural features setting their rates are mostly unknown. Earlier single-molecule fluorescence imaging of the archaeal model glutamate transporter homologue Glt from Pyrococcus horikoshii suggested that the slow conformational transition from the outward- to the inward-facing state, when the bound substrate is translocated from the extracellular to the cytoplasmic side of the membrane, is rate limiting to transport. Here, we provide insight into the structure of the high-energy transition state of Glt that limits the rate of the substrate translocation process. Using bioinformatics, we identified Glt gain-of-function mutations in the flexible helical hairpin domain HP2 and applied linear free energy relationship analysis to infer that the transition state structurally resembles the inward-facing conformation. Based on these analyses, we propose an approach to search for allosteric modulators for transporters.

PubMed: 33185289
DOI: 10.15252/embj.2020105415

Experimental method

X-RAY DIFFRACTION (3.38 Å)