Related ArticlesUse of paramagnetic 19F NMR to monitor domain movement in a glutamate transporter homolog.
Nat Chem Biol. 2020 Jun 08;:
Authors: Huang Y, Wang X, Lv G, Razavi AM, Huysmans GHM, Weinstein H, Bracken C, Eliezer D, Boudker O
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 19F-NMR spectroscopy approach to probe dynamics of large membrane proteins. We labeled a glutamate transporter homolog with a 19F probe via cysteine chemistry and with a Ni2+ ion via chelation by a di-histidine motif. We used distance-dependent enhancement of the longitudinal relaxation of 19F 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 19F nuclei.
PMID: 32514183 [PubMed - as supplied by publisher]
[NMR paper] Utilizing tagged paramagnetic shift reagents to monitor protein dynamics by NMR.
Utilizing tagged paramagnetic shift reagents to monitor protein dynamics by NMR.
Related Articles Utilizing tagged paramagnetic shift reagents to monitor protein dynamics by NMR.
Biochim Biophys Acta. 2017 Sep 22;:
Authors: Ye L, van Eps N, Li X, Ernst OP, Scott Prosser R
Abstract
Calmodulin is a ubiquitous calcium sensor protein, known to serve as a critical interaction hub with a wide range of signaling partners. While the holo form of calmodulin (CaM-4Ca(2+)) has a well-defined ground state structure, it has been shown to...
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Utilizing tagged paramagnetic shift reagents to monitor protein dynamics by NMR
Utilizing tagged paramagnetic shift reagents to monitor protein dynamics by NMR
Publication date: Available online 22 September 2017
Source:Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics</br>
Author(s): Libin Ye, Ned van Eps, Xiang Li, Oliver P. Ernst, R. Scott Prosser</br>
Calmodulin is a ubiquitous calcium sensor protein, known to serve as a critical interaction hub with a wide range of signaling partners. While the holo form of calmodulin (CaM-4Ca2+) has a well-defined ground state structure, it has been shown to undergo exchange, on a...
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09-24-2017 06:16 AM
[NMR paper] In vivo Proton NMR spectroscopy of genetic mouse models BALB/cJ and C57BL/6By: variation in hippocampal glutamate level and the metabotropic glutamate receptor, subtype 7 (Grm7) gene.
In vivo Proton NMR spectroscopy of genetic mouse models BALB/cJ and C57BL/6By: variation in hippocampal glutamate level and the metabotropic glutamate receptor, subtype 7 (Grm7) gene.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles In vivo Proton NMR spectroscopy of genetic mouse models BALB/cJ and C57BL/6By: variation in hippocampal glutamate level and the metabotropic glutamate receptor, subtype 7 (Grm7) gene.
J Mol Neurosci. 2014 May;53(1):135-41
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[NMR paper] Inter-domain dynamics explored by paramagnetic NMR.
Inter-domain dynamics explored by paramagnetic NMR.
Related Articles Inter-domain dynamics explored by paramagnetic NMR.
J Am Chem Soc. 2013 Oct 11;
Authors: Russo L, Maestre-Martínez M, Wolff S, Becker S, Griesinger C
Abstract
An ensemble-based approach is presented to explore the conformational space sampled by a multi-domain protein showing moderate inter-domain dynamics in terms of translational and rotational motions. The strategy was applied on a complex of calmodulin (CaM) with the IQ-recognition motif from the voltage-gated calcium...
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10-12-2013 05:24 PM
[NMR paper] Conformational stabilization of the membrane embedded targeting domain of the lysosomal peptide transporter TAPL for solution NMR.
Conformational stabilization of the membrane embedded targeting domain of the lysosomal peptide transporter TAPL for solution NMR.
Conformational stabilization of the membrane embedded targeting domain of the lysosomal peptide transporter TAPL for solution NMR.
J Biomol NMR. 2013 Sep 7;
Authors: Tumulka F, Roos C, Löhr F, Bock C, Bernhard F, Dötsch V, Abele R
Abstract
The ATP binding cassette transporter TAPL translocates cytosolic peptides into the lumen of lysosomes driven by the hydrolysis of ATP. Functionally, this transporter can be...
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09-10-2013 08:44 PM
[NMR paper] (1)H, (13)C, (15)N backbone and side chain NMR resonance assignments of the N-terminal NEAr iron transporter domain 1 (NEAT 1) of the hemoglobin receptor IsdB of Staphylococcus aureus.
(1)H, (13)C, (15)N backbone and side chain NMR resonance assignments of the N-terminal NEAr iron transporter domain 1 (NEAT 1) of the hemoglobin receptor IsdB of Staphylococcus aureus.
Related Articles (1)H, (13)C, (15)N backbone and side chain NMR resonance assignments of the N-terminal NEAr iron transporter domain 1 (NEAT 1) of the hemoglobin receptor IsdB of Staphylococcus aureus.
Biomol NMR Assign. 2013 May 18;
Authors: Fonner BA, Tripet BP, Lui M, Zhu H, Lei B, Copié V
Abstract
Staphylococcus aureus is an opportunistic pathogen that...
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05-21-2013 02:34 PM
Role of aminotransferases in glutamate metabolism of human erythrocytes
Role of aminotransferases in glutamate metabolism of human erythrocytes
Abstract Human erythrocytes require a continual supply of glutamate to support glutathione synthesis, but are unable to transport this amino acid across their cell membrane. Consequently, erythrocytes rely on de novo glutamate biosynthesis from α-ketoglutarate and glutamine to maintain intracellular levels of glutamate. Erythrocytic glutamate biosynthesis is catalyzed by three enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and glutamine aminohydrolase (GA). Although the presence of these...
Use of paramagnetic 19F NMR to monitor domain movement in a glutamate transporter homolog.
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