hMRAPα, but Not hMRAP2, Enhances hMC4R Constitutive Activity in HEK293 Cells and This Is Not Dependent on hMRAPα Induced Changes in hMC4R Complex N-linked Glycosylation.

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dc.contributor.author Kay, Emma I en
dc.contributor.author Botha, Rikus en
dc.contributor.author Montgomery, Johanna en
dc.contributor.author Mountjoy, Kathleen en
dc.date.accessioned 2020-02-11T03:54:40Z en
dc.date.issued 2015-01 en
dc.identifier.citation PLoS One 10(10):22 pages Article number e0140320 15 Oct 2015 en
dc.identifier.issn 1932-6203 en
dc.identifier.uri http://hdl.handle.net/2292/49891 en
dc.description.abstract MRAP1 but not MRAP2, is essential for melanocortin receptor 2 functional expression. Human MRAP1 splice variant (hMRAPα) and human MRAP2 (hMRAP2) also interact with the other melanocortin receptor subtypes in vitro, although the physiological significance of these interactions is unknown. Previously we showed that HA-hMC4R co-expression with hMRAPα, but not hMRAP2, specifically alters HA-hMC4R complex N-linked glycosylation. hMRAPα-FLAG also enhances hMC4R constitutive activity in vitro. Here we directly compare hMRAPα and hMRAP2 effects on hMC4R constitutive activity in HEK293 cells. In contrast to hMRAPα, co-expression with hMRAP2 had no effect on HA-hMC4R or untagged hMC4R constitutive coupling to adenylyl cyclase. We used fixed and live cell imaging of HA-hMC4R and hMC4R-eGFP respectively, to further characterise effects of hMRAPα on hMC4R subcellular trafficking. hMRAPα-FLAG co-expression did not alter the partitioning of either HA-hMC4R or hMC4R-eGFP into either the ER or the Golgi apparatus, therefore the hMRAPα effect on hMC4R complex N-linked glycosylation is probably not due to hMC4R retention in the ER. We also observed that unlike HA-hMC4R, hMC4R-eGFP lacks complex glycosylation both in the presence and absence of hMRAPα, although both HA-hMC4R and hMC4R-eGFP exhibited increased constitutive coupling to adenylyl cyclase following co-expression with hMRAPα. We conclude that hMRAPα and not hMRAP2 modulates hMC4R constitutive activity. Furthermore, hMRAPα does not increase hMC4R constitutive activity by altering hMC4R complex N-linked glycosylation. Instead we hypothesise that hMRAPα alters hMC4R conformational states leading to increased hMC4R constitutive activity. en
dc.format.medium Electronic-eCollection en
dc.language eng en
dc.relation.ispartofseries PloS one en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.rights.uri https://creativecommons.org/licenses/by/4.0/ en
dc.subject Humans en
dc.subject Carrier Proteins en
dc.subject Membrane Proteins en
dc.subject Receptor, Melanocortin, Type 4 en
dc.subject Protein Isoforms en
dc.subject Alternative Splicing en
dc.subject Protein Conformation en
dc.subject Glycosylation en
dc.subject HEK293 Cells en
dc.subject Adenylyl Cyclases en
dc.title hMRAPα, but Not hMRAP2, Enhances hMC4R Constitutive Activity in HEK293 Cells and This Is Not Dependent on hMRAPα Induced Changes in hMC4R Complex N-linked Glycosylation. en
dc.type Journal Article en
dc.identifier.doi 10.1371/journal.pone.0140320 en
pubs.issue 10 en
pubs.begin-page e0140320 en
pubs.volume 10 en
dc.rights.holder Copyright: The authors en
dc.identifier.pmid 26469516 en
pubs.publication-status Published en
dc.rights.accessrights http://purl.org/eprint/accessRights/OpenAccess en
pubs.subtype research-article en
pubs.subtype Journal Article en
pubs.elements-id 502694 en
pubs.org-id Medical and Health Sciences en
pubs.org-id Medical Sciences en
pubs.org-id Physiology Division en
pubs.org-id Science en
pubs.org-id Science Research en
pubs.org-id Maurice Wilkins Centre (2010-2014) en
dc.identifier.eissn 1932-6203 en
pubs.record-created-at-source-date 2015-10-16 en
pubs.dimensions-id 26469516 en


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