Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility
<div><p>Clustering of ligand:receptor complexes on the cell membrane is widely presumed to have functional consequences for subsequent signal transduction. However, it is experimentally challenging to selectively manipulate receptor clustering without altering other biochemical aspects o...
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| مؤلفون آخرون: | , , , |
| منشور في: |
2021
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| الموضوعات: | |
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| _version_ | 1864513515335712768 |
|---|---|
| author | Zhongwen Chen (770401) |
| author2 | Dongmyung Oh (2361895) Kabir Hassan Biswas (18595561) Ronen Zaidel-Bar (711501) Jay T Groves (18595564) |
| author2_role | author author author author |
| author_facet | Zhongwen Chen (770401) Dongmyung Oh (2361895) Kabir Hassan Biswas (18595561) Ronen Zaidel-Bar (711501) Jay T Groves (18595564) |
| author_role | author |
| dc.creator.none.fl_str_mv | Zhongwen Chen (770401) Dongmyung Oh (2361895) Kabir Hassan Biswas (18595561) Ronen Zaidel-Bar (711501) Jay T Groves (18595564) |
| dc.date.none.fl_str_mv | 2021-08-20T03:00:00Z |
| dc.identifier.none.fl_str_mv | 10.7554/elife.67379 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/Probing_the_effect_of_clustering_on_EphA2_receptor_signaling_efficiency_by_subcellular_control_of_ligand-receptor_mobility/25877974 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biological sciences Biochemistry and cell biology Ligand-receptor complexes Cell membrane Signal transduction Receptor clustering Microfabrication Substrates Mobile ligands |
| dc.title.none.fl_str_mv | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <div><p>Clustering of ligand:receptor complexes on the cell membrane is widely presumed to have functional consequences for subsequent signal transduction. However, it is experimentally challenging to selectively manipulate receptor clustering without altering other biochemical aspects of the cellular system. Here, we develop a microfabrication strategy to produce substrates displaying mobile and immobile ligands that are separated by roughly 1 µm, and thus experience an identical cytoplasmic signaling state, enabling precision comparison of downstream signaling reactions. Applying this approach to characterize the ephrinA1:EphA2 signaling system reveals that EphA2 clustering enhances both receptor phosphorylation and downstream signaling activity. Single-molecule imaging clearly resolves increased molecular binding dwell times at EphA2 clusters for both Grb2:SOS and NCK:N-WASP signaling modules. This type of intracellular comparison enables a substantially higher degree of quantitative analysis than is possible when comparisons must be made between different cells and essentially eliminates the effects of cellular response to ligand manipulation.</p><p> </p></div><h2>Other Information</h2> <p> Published in: eLife<br> License: <a href="http://creativecommons.org/licenses/by/4.0/" target="_blank">http://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.7554/elife.67379" target="_blank">https://dx.doi.org/10.7554/elife.67379</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_242b7534abf02e753a219f488b124ad0 |
| identifier_str_mv | 10.7554/elife.67379 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/25877974 |
| publishDate | 2021 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobilityZhongwen Chen (770401)Dongmyung Oh (2361895)Kabir Hassan Biswas (18595561)Ronen Zaidel-Bar (711501)Jay T Groves (18595564)Biological sciencesBiochemistry and cell biologyLigand-receptor complexesCell membraneSignal transductionReceptor clusteringMicrofabricationSubstratesMobile ligands<div><p>Clustering of ligand:receptor complexes on the cell membrane is widely presumed to have functional consequences for subsequent signal transduction. However, it is experimentally challenging to selectively manipulate receptor clustering without altering other biochemical aspects of the cellular system. Here, we develop a microfabrication strategy to produce substrates displaying mobile and immobile ligands that are separated by roughly 1 µm, and thus experience an identical cytoplasmic signaling state, enabling precision comparison of downstream signaling reactions. Applying this approach to characterize the ephrinA1:EphA2 signaling system reveals that EphA2 clustering enhances both receptor phosphorylation and downstream signaling activity. Single-molecule imaging clearly resolves increased molecular binding dwell times at EphA2 clusters for both Grb2:SOS and NCK:N-WASP signaling modules. This type of intracellular comparison enables a substantially higher degree of quantitative analysis than is possible when comparisons must be made between different cells and essentially eliminates the effects of cellular response to ligand manipulation.</p><p> </p></div><h2>Other Information</h2> <p> Published in: eLife<br> License: <a href="http://creativecommons.org/licenses/by/4.0/" target="_blank">http://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.7554/elife.67379" target="_blank">https://dx.doi.org/10.7554/elife.67379</a></p>2021-08-20T03:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.7554/elife.67379https://figshare.com/articles/journal_contribution/Probing_the_effect_of_clustering_on_EphA2_receptor_signaling_efficiency_by_subcellular_control_of_ligand-receptor_mobility/25877974CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/258779742021-08-20T03:00:00Z |
| spellingShingle | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility Zhongwen Chen (770401) Biological sciences Biochemistry and cell biology Ligand-receptor complexes Cell membrane Signal transduction Receptor clustering Microfabrication Substrates Mobile ligands |
| status_str | publishedVersion |
| title | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| title_full | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| title_fullStr | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| title_full_unstemmed | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| title_short | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| title_sort | Probing the effect of clustering on EphA2 receptor signaling efficiency by subcellular control of ligand-receptor mobility |
| topic | Biological sciences Biochemistry and cell biology Ligand-receptor complexes Cell membrane Signal transduction Receptor clustering Microfabrication Substrates Mobile ligands |