Peptide surfactants for cell-free production of functional G proteincoupled receptors
Two major bottlenecks in elucidating the structure and function of membrane proteins are the difficulty of producing large quantities of functional receptors, and stabilizing them for a sufficient period of time. Selecting the right surfactant is thus crucial. Here we report using peptide surfactant...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
National Academy of Sciences (U.S.)
2011
|
| Online Access: | http://hdl.handle.net/1721.1/67446 |
| _version_ | 1811072565774385152 |
|---|---|
| author | Wang, Xiaoqiang Corin, Karolina A. Baaske, Philipp Wienken, Christoph J. Jerabek-Willemsen, Moran Duhr, Stefan Braun, Dieter Zhang, Shuguang |
| author2 | Massachusetts Institute of Technology. Center for Biomedical Engineering |
| author_facet | Massachusetts Institute of Technology. Center for Biomedical Engineering Wang, Xiaoqiang Corin, Karolina A. Baaske, Philipp Wienken, Christoph J. Jerabek-Willemsen, Moran Duhr, Stefan Braun, Dieter Zhang, Shuguang |
| author_sort | Wang, Xiaoqiang |
| collection | MIT |
| description | Two major bottlenecks in elucidating the structure and function of membrane proteins are the difficulty of producing large quantities of functional receptors, and stabilizing them for a sufficient period of time. Selecting the right surfactant is thus crucial. Here we report using peptide surfactants in commercial Escherichia coli cell-free systems to rapidly produce milligram quantities of soluble G protein-coupled receptors (GPCRs). These include the human formyl peptide receptor, human trace amine-associated receptor, and two olfactory receptors. The GPCRs expressed in the presence of the peptide surfactants were soluble and had α-helical secondary structures, suggesting that they were properly folded. Microscale thermophoresis measurements showed that one olfactory receptor expressed using peptide surfactants bound its known ligand heptanal (molecular weight 114.18). These short and simple peptide surfactants may be able to facilitate the rapid production of GPCRs, or even other membrane proteins, for structure and function studies. |
| first_indexed | 2024-09-23T09:07:57Z |
| format | Article |
| id | mit-1721.1/67446 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:07:57Z |
| publishDate | 2011 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/674462022-09-30T13:38:13Z Peptide surfactants for cell-free production of functional G proteincoupled receptors Wang, Xiaoqiang Corin, Karolina A. Baaske, Philipp Wienken, Christoph J. Jerabek-Willemsen, Moran Duhr, Stefan Braun, Dieter Zhang, Shuguang Massachusetts Institute of Technology. Center for Biomedical Engineering Zhang, Shuguang Zhang, Shuguang Wang, Xiaoqiang Corin, Karolina A. Two major bottlenecks in elucidating the structure and function of membrane proteins are the difficulty of producing large quantities of functional receptors, and stabilizing them for a sufficient period of time. Selecting the right surfactant is thus crucial. Here we report using peptide surfactants in commercial Escherichia coli cell-free systems to rapidly produce milligram quantities of soluble G protein-coupled receptors (GPCRs). These include the human formyl peptide receptor, human trace amine-associated receptor, and two olfactory receptors. The GPCRs expressed in the presence of the peptide surfactants were soluble and had α-helical secondary structures, suggesting that they were properly folded. Microscale thermophoresis measurements showed that one olfactory receptor expressed using peptide surfactants bound its known ligand heptanal (molecular weight 114.18). These short and simple peptide surfactants may be able to facilitate the rapid production of GPCRs, or even other membrane proteins, for structure and function studies. United States. Defense Advanced Research Projects Agency (HR0011-09-C-0012) China University of Petroleum NanoSystems Initiative Munich Universität München (Center for NanoScience) 2011-12-05T21:20:18Z 2011-12-05T21:20:18Z 2011-05 2010-12 Article http://purl.org/eprint/type/JournalArticle 1091-6490 http://hdl.handle.net/1721.1/67446 Wang, X. et al. “Peptide surfactants for cell-free production of functional G protein-coupled receptors.” Proceedings of the National Academy of Sciences 108.22 (2011): 9049-9054. ©2011 by the National Academy of Sciences. en_US http://dx.doi.org/10.1073/pnas.1018185108 Proceedings of the National Academy of Sciences of the United States of America Article 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. application/pdf National Academy of Sciences (U.S.) PNAS |
| spellingShingle | Wang, Xiaoqiang Corin, Karolina A. Baaske, Philipp Wienken, Christoph J. Jerabek-Willemsen, Moran Duhr, Stefan Braun, Dieter Zhang, Shuguang Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title | Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title_full | Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title_fullStr | Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title_full_unstemmed | Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title_short | Peptide surfactants for cell-free production of functional G proteincoupled receptors |
| title_sort | peptide surfactants for cell free production of functional g proteincoupled receptors |
| url | http://hdl.handle.net/1721.1/67446 |
| work_keys_str_mv | AT wangxiaoqiang peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT corinkarolinaa peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT baaskephilipp peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT wienkenchristophj peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT jerabekwillemsenmoran peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT duhrstefan peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT braundieter peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors AT zhangshuguang peptidesurfactantsforcellfreeproductionoffunctionalgproteincoupledreceptors |