7ngh: Difference between revisions
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==Structure of glutamate transporter homologue in complex with Sybody== | |||
<StructureSection load='7ngh' size='340' side='right'caption='[[7ngh]], [[Resolution|resolution]] 3.50Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[7ngh]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Thermococcus_kodakarensis_KOD1 Thermococcus kodakarensis KOD1] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7NGH OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7NGH FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 3.5Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ASP:ASPARTIC+ACID'>ASP</scene></td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7ngh FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7ngh OCA], [https://pdbe.org/7ngh PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7ngh RCSB], [https://www.ebi.ac.uk/pdbsum/7ngh PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7ngh ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
It is well-established that the secondary active transporters Glt(Tk) and Glt(Ph) catalyze coupled uptake of aspartate and three sodium ions, but insight in the kinetic mechanism of transport is fragmentary. Here, we systematically measured aspartate uptake rates in proteoliposomes containing purified Glt(Tk), and derived the rate equation for a mechanism in which two sodium ions bind before and another after aspartate. Re-analysis of existing data on Glt(Ph) using this equation allowed for determination of the turnover number (0.14 s(-1)), without the need for error-prone protein quantification. To overcome the complication that purified transporters may adopt right-side-out or inside-out membrane orientations upon reconstitution, thereby confounding the kinetic analysis, we employed a rapid method using synthetic nanobodies to inactivate one population. Oppositely oriented Glt(Tk) proteins showed the same transport kinetics, consistent with the use of an identical gating element on both sides of the membrane. Our work underlines the value of bona fide transport experiments to reveal mechanistic features of Na(+)-aspartate symport that cannot be observed in detergent solution. Combined with previous pre-equilibrium binding studies, a full kinetic mechanism of structurally characterized aspartate transporters of the SLC1A family is now emerging. | |||
Kinetic mechanism of Na(+)-coupled aspartate transport catalyzed by Glt(Tk).,Trinco G, Arkhipova V, Garaeva AA, Hutter CAJ, Seeger MA, Guskov A, Slotboom DJ Commun Biol. 2021 Jun 17;4(1):751. doi: 10.1038/s42003-021-02267-y. PMID:34140623<ref>PMID:34140623</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 7ngh" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Symporter 3D structures|Symporter 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Synthetic construct]] | |||
[[Category: Thermococcus kodakarensis KOD1]] | |||
[[Category: Arkhipova V]] | |||
[[Category: Guskov A]] | |||
[[Category: Slotboom DJ]] | |||
Latest revision as of 12:30, 9 October 2024
Structure of glutamate transporter homologue in complex with SybodyStructure of glutamate transporter homologue in complex with Sybody
Structural highlights
Publication Abstract from PubMedIt is well-established that the secondary active transporters Glt(Tk) and Glt(Ph) catalyze coupled uptake of aspartate and three sodium ions, but insight in the kinetic mechanism of transport is fragmentary. Here, we systematically measured aspartate uptake rates in proteoliposomes containing purified Glt(Tk), and derived the rate equation for a mechanism in which two sodium ions bind before and another after aspartate. Re-analysis of existing data on Glt(Ph) using this equation allowed for determination of the turnover number (0.14 s(-1)), without the need for error-prone protein quantification. To overcome the complication that purified transporters may adopt right-side-out or inside-out membrane orientations upon reconstitution, thereby confounding the kinetic analysis, we employed a rapid method using synthetic nanobodies to inactivate one population. Oppositely oriented Glt(Tk) proteins showed the same transport kinetics, consistent with the use of an identical gating element on both sides of the membrane. Our work underlines the value of bona fide transport experiments to reveal mechanistic features of Na(+)-aspartate symport that cannot be observed in detergent solution. Combined with previous pre-equilibrium binding studies, a full kinetic mechanism of structurally characterized aspartate transporters of the SLC1A family is now emerging. Kinetic mechanism of Na(+)-coupled aspartate transport catalyzed by Glt(Tk).,Trinco G, Arkhipova V, Garaeva AA, Hutter CAJ, Seeger MA, Guskov A, Slotboom DJ Commun Biol. 2021 Jun 17;4(1):751. doi: 10.1038/s42003-021-02267-y. PMID:34140623[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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