A Computationally Designed Water-Soluble Variant of a G-Protein-Coupled Receptor: The Human Mu Opioid Receptor

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dc.contributor.authorPerez Aguilar, Jose Manuel
dc.contributor.authorXi, Jin
dc.contributor.authorMatsunaga, Felipe
dc.contributor.authorCui, Xu
dc.contributor.authorSaven, Jeffery G.
dc.contributor.authorSelling, Bernard
dc.contributor.authorLiu, Renyu
dc.date2023-05-17T14:36:49.000
dc.date.accessioned2023-05-22T12:45:42Z
dc.date.available2023-05-22T12:45:42Z
dc.date.issued2013-06-14
dc.date.submitted2016-06-12T17:17:19-07:00
dc.description.abstractG-protein-coupled receptors (GPCRs) play essential roles in various physiological processes, and are widely targeted by pharmaceutical drugs. Despite their importance, studying GPCRs has been problematic due to difficulties in isolating large quantities of these membrane proteins in forms that retain their ligand binding capabilities. Creating water-soluble variants of GPCRs by mutating the exterior, transmembrane residues provides a potential method to overcome these difficulties. Here we present the first study involving the computational design, expression and characterization of water-soluble variant of a human GPCR, the human mu opioid receptor (MUR), which is involved in pain and addiction. An atomistic structure of the transmembrane domain was built using comparative (homology) modeling and known GPCR structures. This structure was highly similar to the subsequently determined structure of the murine receptor and was used to computationally design 53 mutations of exterior residues in the transmembrane region, yielding a variant intended to be soluble in aqueous media. The designed variant expressed in high yield in Escherichia coli and was water soluble. The variant shared structural and functionally related features with the native human MUR, including helical secondary structure and comparable affinity for the antagonist naltrexone (Kd  = 65 nM). The roles of cholesterol and disulfide bonds on the stability of the receptor variant were also investigated. This study exemplifies the potential of the computational approach to produce water-soluble variants of GPCRs amenable for structural and functionally related characterization in aqueous solution.
dc.identifier.urihttps://repository.upenn.edu/handle/20.500.14332/6101
dc.legacy.articleid1001
dc.legacy.fields10.1371/journal.pone.0066009
dc.legacy.fulltexturlhttps://repository.upenn.edu/cgi/viewcontent.cgi?article=1001&context=chemistry_papers&unstamped=1
dc.rights© 2013 Perez-Aguilar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
dc.source.beginpagee66009-1
dc.source.endpagee66009-10
dc.source.issue24
dc.source.issue6
dc.source.journalDepartmental Papers (Chemistry)
dc.source.journaltitlePLoS ONE
dc.source.peerreviewedtrue
dc.source.statuspublished
dc.source.volume8
dc.subject.otherBiochemistry
dc.subject.otherOrganic Chemistry
dc.subject.otherPhysical Chemistry
dc.titleA Computationally Designed Water-Soluble Variant of a G-Protein-Coupled Receptor: The Human Mu Opioid Receptor
dc.typeArticle
digcom.contributor.authorisAuthorOfPublication|email:perezagu@sas.upenn.edu|institution:University of Pennsylvania|Perez Aguilar, Jose Manuel
digcom.contributor.authorXi, Jin
digcom.contributor.authorMatsunaga, Felipe
digcom.contributor.authorCui, Xu
digcom.contributor.authorSelling, Bernard
digcom.contributor.authorisAuthorOfPublication|email:saven@sas.upenn.edu|institution:University of Pennsylvania|Saven, Jeffery G.
digcom.contributor.authorLiu, Renyu
digcom.identifierchemistry_papers/24
digcom.identifier.contextkey8719265
digcom.identifier.submissionpathchemistry_papers/24
digcom.typearticle
dspace.entity.typePublication
relation.isAuthorOfPublication534f9097-0322-4127-9146-b784c78e4b64
relation.isAuthorOfPublication4951796f-63aa-4a0f-a9d3-ae7d800a4c50
relation.isAuthorOfPublication4951796f-63aa-4a0f-a9d3-ae7d800a4c50
relation.isAuthorOfPublication.latestForDiscovery534f9097-0322-4127-9146-b784c78e4b64
upenn.schoolDepartmentCenterDepartmental Papers (Chemistry)
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