Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering

<p dir="ltr">A multitude of membrane-localized receptors are utilized by cells to integrate both biochemical and physical signals from their microenvironment. The clustering of membrane receptors is widely presumed to have functional consequences for subsequent signal transduction. H...

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Main Author: Zhongwen Chen (770401) (author)
Other Authors: Kabir Biswas (18278695) (author), Jay Groves (520696) (author)
Published: 2022
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author Zhongwen Chen (770401)
author2 Kabir Biswas (18278695)
Jay Groves (520696)
author2_role author
author
author_facet Zhongwen Chen (770401)
Kabir Biswas (18278695)
Jay Groves (520696)
author_role author
dc.creator.none.fl_str_mv Zhongwen Chen (770401)
Kabir Biswas (18278695)
Jay Groves (520696)
dc.date.none.fl_str_mv 2022-06-05T06:00:00Z
dc.identifier.none.fl_str_mv 10.21769/bioprotoc.4434
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Patterned_Substrate_of_Mobile_and_Immobile_Ligands_to_Probe_EphA2_Receptor_Clustering/25513876
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biological sciences
Genetics
Biomedical and clinical sciences
Neurosciences
Supported lipid bilayer
Micropatterning
EphA2 receptor
Receptor clustering
Signaling transduction
dc.title.none.fl_str_mv Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <p dir="ltr">A multitude of membrane-localized receptors are utilized by cells to integrate both biochemical and physical signals from their microenvironment. The clustering of membrane receptors 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 describe a method to fabricate multicomponent, ligand-functionalized microarrays, for spatially segregated and simultaneous monitoring of receptor activation and signaling in individual living cells. While existing micropatterning techniques allow for the display of fixed ligands, this protocol uniquely allows for functionalization of both mobile membrane corrals and immobile polymers with selective ligands, as well as microscopic monitoring of cognate receptor activation at the cell membrane interface. This protocol has been developed to study the effects of clustering on EphA2 signaling transduction. It is potentially applicable to multiple cell signaling systems, or microbe/host interactions.</p><h2>Other Information</h2><p dir="ltr">Published in: BIO-PROTOCOL<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.21769/bioprotoc.4434" target="_blank">https://dx.doi.org/10.21769/bioprotoc.4434</a></p>
eu_rights_str_mv openAccess
id Manara2_8bf50221d2b55a99148f400ef10e8285
identifier_str_mv 10.21769/bioprotoc.4434
network_acronym_str Manara2
network_name_str Manara2
oai_identifier_str oai:figshare.com:article/25513876
publishDate 2022
repository.mail.fl_str_mv
repository.name.fl_str_mv
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rights_invalid_str_mv CC BY 4.0
spelling Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor ClusteringZhongwen Chen (770401)Kabir Biswas (18278695)Jay Groves (520696)Biological sciencesGeneticsBiomedical and clinical sciencesNeurosciencesSupported lipid bilayerMicropatterningEphA2 receptorReceptor clusteringSignaling transduction<p dir="ltr">A multitude of membrane-localized receptors are utilized by cells to integrate both biochemical and physical signals from their microenvironment. The clustering of membrane receptors 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 describe a method to fabricate multicomponent, ligand-functionalized microarrays, for spatially segregated and simultaneous monitoring of receptor activation and signaling in individual living cells. While existing micropatterning techniques allow for the display of fixed ligands, this protocol uniquely allows for functionalization of both mobile membrane corrals and immobile polymers with selective ligands, as well as microscopic monitoring of cognate receptor activation at the cell membrane interface. This protocol has been developed to study the effects of clustering on EphA2 signaling transduction. It is potentially applicable to multiple cell signaling systems, or microbe/host interactions.</p><h2>Other Information</h2><p dir="ltr">Published in: BIO-PROTOCOL<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.21769/bioprotoc.4434" target="_blank">https://dx.doi.org/10.21769/bioprotoc.4434</a></p>2022-06-05T06:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.21769/bioprotoc.4434https://figshare.com/articles/journal_contribution/Patterned_Substrate_of_Mobile_and_Immobile_Ligands_to_Probe_EphA2_Receptor_Clustering/25513876CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/255138762022-06-05T06:00:00Z
spellingShingle Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
Zhongwen Chen (770401)
Biological sciences
Genetics
Biomedical and clinical sciences
Neurosciences
Supported lipid bilayer
Micropatterning
EphA2 receptor
Receptor clustering
Signaling transduction
status_str publishedVersion
title Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
title_full Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
title_fullStr Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
title_full_unstemmed Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
title_short Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
title_sort Patterned Substrate of Mobile and Immobile Ligands to Probe EphA2 Receptor Clustering
topic Biological sciences
Genetics
Biomedical and clinical sciences
Neurosciences
Supported lipid bilayer
Micropatterning
EphA2 receptor
Receptor clustering
Signaling transduction