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Hall, Jenny L., Sohail, Azmat, Cabrita, Eurico J., Macdonald, Colin, Stockner, Thomas, Sitte, Harald H., Angulo, Jesus 
ORCID: https://orcid.org/0000-0001-7250-5639 and Macmillan, Fraser ORCID: https://orcid.org/0000-0002-2410-4790
(2020)
Saturation transfer difference NMR on the integral trimeric membrane transport protein GltPh determines cooperative substrate binding.
Scientific Reports, 10.
ISSN 2045-2322
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Abstract
Saturation-transfer difference (STD) NMR spectroscopy is a fast and versatile method which can be applied for drug-screening purposes, allowing the determination of essential ligand binding affinities (KD). Although widely employed to study soluble proteins, its use remains negligible for membrane proteins. Here the use of STD NMR for KD determination is demonstrated for two competing substrates with very different binding affinities (low nanomolar to millimolar) for an integral membrane transport protein in both detergent-solubilised micelles and reconstituted proteoliposomes. GltPh, a homotrimeric aspartate transporter from Pyrococcus horikoshii, is an archaeal homolog of mammalian membrane transport proteins—known as excitatory amino acid transporters (EAATs). They are found within the central nervous system and are responsible for fast uptake of the neurotransmitter glutamate, essential for neuronal function. Differences in both KD’s and cooperativity are observed between detergent micelles and proteoliposomes, the physiological implications of which are discussed.
| Item Type: | Article |
|---|---|
| Faculty \ School: | Faculty of Science > School of Chemistry Faculty of Science Faculty of Science > School of Pharmacy |
| UEA Research Groups: | Faculty of Science > Research Groups > Pharmaceutical Materials and Soft Matter Faculty of Science > Research Groups > Chemistry of Life Processes Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Chemistry of Light and Energy |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 13 Oct 2020 00:05 |
| Last Modified: | 31 Jan 2024 02:47 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/77224 |
| DOI: | 10.1038/s41598-020-73443-z |
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