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dc.contributor.authorYang, Heehong
dc.contributor.authorSong, Hyun Seok
dc.contributor.authorAhn, Sae Ryun
dc.contributor.authorPark, Tai Hyun
dc.date.accessioned2017-01-26T15:51:07Z
dc.date.available2017-01-26T15:51:07Z
dc.date.issued2015-07
dc.date.submitted2015-03
dc.identifier.issn1226-8372
dc.identifier.issn1976-3816
dc.identifier.urihttp://hdl.handle.net/1721.1/106634
dc.description.abstractOlfactory receptors (ORs), belonging to the Gprotein coupled receptor (GPCR) family, are very difficult to be overexpressed, purified and reconstituted because of their hydrophobicity and complicated structure. These receptors bind to their specific ligands, thus their specificity is very useful for application as a bioelectronic nose. Furthermore, highly purified and well-reconstituted human olfactory receptor (hOR) can be used in various fields, such as in protein-interaction research, drug screening, and analysis of the hOR structure. In this study, human olfactory receptor, hOR2AG1, was produced with high purity and functionally reconstituted in detergent micelles. The hOR2AG1 was overexpressed in Escherichia coli (E. coli) with glutathione S-transferase (GST) and 6xHis-tag as an inclusion body. The hOR2AG1 fusion protein was solubilized in buffer containing sodium dodecyl sulfate (SDS) and purified using Ni-NTA chromatography. The GST domain was removed using proteolytic cleavage before elution from the column. After purification, the hOR2AG1 was successfully reconstituted using nonionic detergents and methyl-ß-cyclodextrin. Finally highly purified and well-reconstituted hOR was obtained, and its biological characteristics were confirmed by using circular dichroism (CD) spectrum and tryptophan fluorescence assay. These results can be applied to develop protein-based sensing systems including a bioelectronic nose and to analyze the native hOR structure using solid-state NMR, X-ray crystallography, or neutron scattering.en_US
dc.description.sponsorshipKorea (South). Ministry of Science, ICT and Future Planning (Grants2014039771 and 2014053108)en_US
dc.description.sponsorshipKorea Institute of Science and Technology (KIST) (Project 2E24812-14-043)en_US
dc.publisherThe Korean Society for Biotechnology and Bioengineeringen_US
dc.relation.isversionofhttp://dx.doi.org/10.1007/s12257-014-0897-4en_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourceThe Korean Society for Biotechnology and Bioengineeringen_US
dc.titlePurification and functional reconstitution of human olfactory receptor expressed in Escherichia colien_US
dc.typeArticleen_US
dc.identifier.citationYang, Heehong et al. “Purification and Functional Reconstitution of Human Olfactory Receptor Expressed in Escherichia Coli.” Biotechnology and Bioprocess Engineering 20.3 (2015): 423–430.en_US
dc.contributor.departmentHarvard University--MIT Division of Health Sciences and Technologyen_US
dc.contributor.mitauthorSong, Hyun Seok
dc.relation.journalBiotechnology and Bioprocess Engineeringen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2016-08-18T15:47:25Z
dc.language.rfc3066en
dc.rights.holderThe Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg
dspace.orderedauthorsYang, Heehong; Song, Hyun Seok; Ahn, Sae Ryun; Park, Tai Hyunen_US
dspace.embargo.termsNen
mit.licensePUBLISHER_POLICYen_US
mit.metadata.statusComplete


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