dc.contributor.author |
Hunter, Morag R |
en |
dc.contributor.author |
Finlay, David |
en |
dc.contributor.author |
Macdonald, Christa E |
en |
dc.contributor.author |
Cawston, Erin |
en |
dc.contributor.author |
Grimsey, Natasha |
en |
dc.contributor.author |
Glass, Michelle |
en |
dc.date.accessioned |
2018-11-07T21:49:05Z |
en |
dc.date.issued |
2017-01 |
en |
dc.identifier.issn |
0076-6879 |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/44089 |
en |
dc.description.abstract |
Cannabinoid receptors, like other GPCRs, signal via a spectrum of related signaling pathways. Recently, monitoring GPCR-mediated cAMP signaling has become significantly easier with the development of genetically encoded, transfectable cAMP biosensors. Cell lines transfected with these biosensors can be monitored continuously, allowing the analysis of receptor-mediated signaling in unprecedented detail. Here, we describe a protocol for transfectable biosensors which report cellular cAMP concentrations by bioluminescence resonance energy transfer (BRET). This assay system has been utilized to elucidate the temporal nature of agonists and allosteric modulators of the cannabinoid receptor CB1. In particular, the CB1 allosteric modulator ORG27569 has been shown to modify receptor agonism in a time-dependent fashion; a characteristic which would not have been observed via traditional endpoint methods of detecting cAMP signaling. BRET cAMP biosensors are suitable for miniaturization and automation, and as such are valuable and cost-effective tools for moderate- to high-throughput experimental protocols. |
en |
dc.format.medium |
Print-Electronic |
en |
dc.language |
eng |
en |
dc.relation.ispartofseries |
Methods in enzymology |
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.subject |
Humans |
en |
dc.subject |
Receptors, Cannabinoid |
en |
dc.subject |
Cyclic AMP |
en |
dc.subject |
Microscopy, Fluorescence |
en |
dc.subject |
Fluorescence Resonance Energy Transfer |
en |
dc.subject |
Biosensing Techniques |
en |
dc.subject |
Second Messenger Systems |
en |
dc.subject |
Protein Binding |
en |
dc.subject |
HEK293 Cells |
en |
dc.subject |
Colforsin |
en |
dc.subject |
Adenylyl Cyclase Inhibitors |
en |
dc.title |
Real-Time Measurement of Cannabinoid Receptor-Mediated cAMP Signaling. |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1016/bs.mie.2017.05.001 |
en |
pubs.begin-page |
43 |
en |
pubs.volume |
593 |
en |
dc.rights.holder |
Copyright: The author |
en |
dc.identifier.pmid |
28750814 |
en |
pubs.end-page |
59 |
en |
pubs.publication-status |
Published |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Journal Article |
en |
pubs.elements-id |
638575 |
en |
pubs.org-id |
Medical and Health Sciences |
en |
pubs.org-id |
Medical Sciences |
en |
pubs.org-id |
Pharmacology |
en |
dc.identifier.eissn |
1557-7988 |
en |
pubs.record-created-at-source-date |
2017-07-29 |
en |
pubs.dimensions-id |
28750814 |
en |