7r63
From Proteopedia
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==Nb82, a nanobody against voltage gated sodium channels Nav1.4 and Nav1.5== | ==Nb82, a nanobody against voltage gated sodium channels Nav1.4 and Nav1.5== | ||
| - | <StructureSection load='7r63' size='340' side='right'caption='[[7r63]]' scene=''> | + | <StructureSection load='7r63' size='340' side='right'caption='[[7r63]], [[Resolution|resolution]] 2.00Å' scene=''> |
== Structural highlights == | == Structural highlights == | ||
| - | <table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7R63 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7R63 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[7r63]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Lama_glama Lama glama]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7R63 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7R63 FirstGlance]. <br> |
| - | </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=7r63 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7r63 OCA], [https://pdbe.org/7r63 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7r63 RCSB], [https://www.ebi.ac.uk/pdbsum/7r63 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7r63 ProSAT]</span></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2Å</td></tr> |
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=PG0:2-(2-METHOXYETHOXY)ETHANOL'>PG0</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</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=7r63 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7r63 OCA], [https://pdbe.org/7r63 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7r63 RCSB], [https://www.ebi.ac.uk/pdbsum/7r63 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7r63 ProSAT]</span></td></tr> | ||
</table> | </table> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Voltage-gated sodium channels, NaVs, are responsible for the rapid rise of action potentials in excitable tissues. NaV channel mutations have been implicated in several human genetic diseases, such as hypokalemic periodic paralysis, myotonia, and long-QT and Brugada syndromes. Here, we generated high-affinity anti-NaV nanobodies (Nbs), Nb17 and Nb82, that recognize the NaV1.4 (skeletal muscle) and NaV1.5 (cardiac muscle) channel isoforms. These Nbs were raised in llama (Lama glama) and selected from a phage display library for high affinity to the C-terminal (CT) region of NaV1.4. The Nbs were expressed in Escherichia coli, purified, and biophysically characterized. Development of high-affinity Nbs specifically targeting a given human NaV isoform has been challenging because they usually show undesired crossreactivity for different NaV isoforms. Our results show, however, that Nb17 and Nb82 recognize the CTNaV1.4 or CTNaV1.5 over other CTNav isoforms. Kinetic experiments by biolayer interferometry determined that Nb17 and Nb82 bind to the CTNaV1.4 and CTNaV1.5 with high affinity (KD approximately 40-60 nM). In addition, as proof of concept, we show that Nb82 could detect NaV1.4 and NaV1.5 channels in mammalian cells and tissues by Western blot. Furthermore, human embryonic kidney cells expressing holo NaV1.5 channels demonstrated a robust FRET-binding efficiency for Nb17 and Nb82. Our work lays the foundation for developing Nbs as anti-NaV reagents to capture NaVs from cell lysates and as molecular visualization agents for NaVs. | ||
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| + | Development of high-affinity nanobodies specific for NaV1.4 and NaV1.5 voltage-gated sodium channel isoforms.,Srinivasan L, Alzogaray V, Selvakumar D, Nathan S, Yoder JB, Wright KM, Klinke S, Nwafor JN, Labanda MS, Goldbaum FA, Schon A, Freire E, Tomaselli GF, Amzel LM, Ben-Johny M, Gabelli SB J Biol Chem. 2022 Apr;298(4):101763. doi: 10.1016/j.jbc.2022.101763. Epub 2022, Feb 21. PMID:35202650<ref>PMID:35202650</ref> | ||
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| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 7r63" style="background-color:#fffaf0;"></div> | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
| + | [[Category: Lama glama]] | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Gabelli SB]] | [[Category: Gabelli SB]] | ||
[[Category: Srinivasan L]] | [[Category: Srinivasan L]] | ||
Revision as of 16:26, 18 October 2023
Nb82, a nanobody against voltage gated sodium channels Nav1.4 and Nav1.5
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