Show simple item record

dc.contributor.authorShilling, PJ
dc.contributor.authorBumbak, F
dc.contributor.authorScott, DJ
dc.contributor.authorBathgate, RAD
dc.contributor.authorGooley, PR
dc.date.accessioned2021-02-04T00:54:57Z
dc.date.available2021-02-04T00:54:57Z
dc.date.issued2017-04-24
dc.identifierpii: 10.1038/s41598-017-01227-z
dc.identifier.citationShilling, P. J., Bumbak, F., Scott, D. J., Bathgate, R. A. D. & Gooley, P. R. (2017). Characterisation of a cell-free synthesised G-protein coupled receptor. SCIENTIFIC REPORTS, 7 (1), https://doi.org/10.1038/s41598-017-01227-z.
dc.identifier.issn2045-2322
dc.identifier.urihttp://hdl.handle.net/11343/259221
dc.description.abstractG-protein coupled receptors are the largest family of integral membrane proteins found within the human genome. They function as receptors and modulators to a wide range of ligands and responses which are crucial for human health. GPCR study, specifically the investigation of structure and interaction to cognate ligands, is of high priority. Limitations for structural study can be traced in part, to obtaining suitable quantities of recombinant protein. We sought to address the limitations of traditional recombinant technologies by utilising an Escherichia coli based cell-free protein synthesis (CFPS) approach for production of a thermostable neurotensin receptor 1 (en2NTS1). Initial results were promising, with a high amount (up to 2 mg/mL) of en2NTS1 produced, that had attained correct secondary structure. Meanwhile, concurrent experiments indicated that CFPS produced en2NTS1 showed non-competitive binding to the peptide ligand neurotensin8-13 when compared to E. coli produced en2NTS1. 1H-13C HMQC SOFAST NMR spectra were indicative of disrupted tertiary structure for CFPS produced 13CH3-methionine labelled en2NTS1. The results obtained, indicate CFPS produced en2NTS1 is not forming a discrete tertiary structure and that further development of the CFPS technique needs to be carried out.
dc.languageEnglish
dc.publisherNATURE PUBLISHING GROUP
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.titleCharacterisation of a cell-free synthesised G-protein coupled receptor
dc.typeJournal Article
dc.identifier.doi10.1038/s41598-017-01227-z
melbourne.affiliation.departmentFlorey Department of Neuroscience and Mental Health
melbourne.affiliation.departmentBiochemistry and Molecular Biology
melbourne.affiliation.departmentSchool of Chemistry
melbourne.affiliation.facultyMedicine, Dentistry & Health Sciences
melbourne.affiliation.facultyScience
melbourne.source.titleScientific Reports
melbourne.source.volume7
melbourne.source.issue1
melbourne.identifier.nhmrc1081844
dc.rights.licenseCC BY
melbourne.elementsid1205481
melbourne.contributor.authorBumbak, Fabian
melbourne.contributor.authorScott, Daniel
melbourne.contributor.authorBathgate, Ross
melbourne.contributor.authorGooley, Paul
melbourne.contributor.authorShilling, Patrick
dc.identifier.eissn2045-2322
melbourne.identifier.fundernameidNHMRC, 1081844
melbourne.accessrightsOpen Access


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record