Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes
Escherichia coli is a convenient host for the expression of proteins, but the heterologous production of large membrane protein complexes often is hampered by the lack of specific accessory genes required for membrane insertion or cofactor assembly. In this study we introduce the non-pathogenic and...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.02537/full |
| _version_ | 1819054876060024832 |
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| author | Lena Schleicher Valentin Muras Björn Claussen Jens Pfannstiel Bastian Blombach Pavel Dibrov Günter Fritz Günter Fritz Julia Steuber |
| author_facet | Lena Schleicher Valentin Muras Björn Claussen Jens Pfannstiel Bastian Blombach Pavel Dibrov Günter Fritz Günter Fritz Julia Steuber |
| author_sort | Lena Schleicher |
| collection | DOAJ |
| description | Escherichia coli is a convenient host for the expression of proteins, but the heterologous production of large membrane protein complexes often is hampered by the lack of specific accessory genes required for membrane insertion or cofactor assembly. In this study we introduce the non-pathogenic and fast-growing Vibrio natriegens as a suitable expression host for membrane-bound proteins from Vibrio cholerae. We achieved production of the primary Na+ pump, the NADH:quinone oxidoreductase (NQR), from V. cholerae in an active state, as indicated by increased overall NADH:quinone oxidoreduction activity of membranes from the transformed V. natriegens, and the sensitivity toward Ag+, a specific inhibitor of the NQR. Complete assembly of V. cholerae NQR expressed in V. natriegens was demonstrated by BN PAGE followed by activity staining. The secondary transport system Mrp from V. cholerae, another membrane-bound multisubunit complex, was also produced in V. natriegens in a functional state, as demonstrated by in vivo Li+ transport. V. natriegens is a promising expression host for the production of membrane protein complexes from Gram-negative pathogens. |
| first_indexed | 2024-12-21T12:58:35Z |
| format | Article |
| id | doaj.art-16bdcd40b5a2492fb9201bbb741b2c99 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-21T12:58:35Z |
| publishDate | 2018-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-16bdcd40b5a2492fb9201bbb741b2c992022-12-21T19:03:15ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-10-01910.3389/fmicb.2018.02537414211Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein ComplexesLena Schleicher0Valentin Muras1Björn Claussen2Jens Pfannstiel3Bastian Blombach4Pavel Dibrov5Günter Fritz6Günter Fritz7Julia Steuber8Institute of Microbiology, University of Hohenheim, Stuttgart, GermanyInstitute of Microbiology, University of Hohenheim, Stuttgart, GermanyInstitute of Microbiology, University of Hohenheim, Stuttgart, GermanyMass Spectrometry Core Facility, University of Hohenheim, Stuttgart, GermanyInstitute of Biochemical Engineering, University of Stuttgart, Stuttgart, GermanyDepartment of Microbiology, University of Manitoba, Winnipeg, MB, CanadaInstitute of Microbiology, University of Hohenheim, Stuttgart, GermanyInstitute for Neuropathology, University of Freiburg, Freiburg im Breisgau, GermanyInstitute of Microbiology, University of Hohenheim, Stuttgart, GermanyEscherichia coli is a convenient host for the expression of proteins, but the heterologous production of large membrane protein complexes often is hampered by the lack of specific accessory genes required for membrane insertion or cofactor assembly. In this study we introduce the non-pathogenic and fast-growing Vibrio natriegens as a suitable expression host for membrane-bound proteins from Vibrio cholerae. We achieved production of the primary Na+ pump, the NADH:quinone oxidoreductase (NQR), from V. cholerae in an active state, as indicated by increased overall NADH:quinone oxidoreduction activity of membranes from the transformed V. natriegens, and the sensitivity toward Ag+, a specific inhibitor of the NQR. Complete assembly of V. cholerae NQR expressed in V. natriegens was demonstrated by BN PAGE followed by activity staining. The secondary transport system Mrp from V. cholerae, another membrane-bound multisubunit complex, was also produced in V. natriegens in a functional state, as demonstrated by in vivo Li+ transport. V. natriegens is a promising expression host for the production of membrane protein complexes from Gram-negative pathogens.https://www.frontiersin.org/article/10.3389/fmicb.2018.02537/fullVibrio natriegensmembrane proteinsmultisubunit complexesexpressionNa+-translocating NADH:quinone oxidoreductasemultiple resistance and pH related antiporter |
| spellingShingle | Lena Schleicher Valentin Muras Björn Claussen Jens Pfannstiel Bastian Blombach Pavel Dibrov Günter Fritz Günter Fritz Julia Steuber Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes Frontiers in Microbiology Vibrio natriegens membrane proteins multisubunit complexes expression Na+-translocating NADH:quinone oxidoreductase multiple resistance and pH related antiporter |
| title | Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes |
| title_full | Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes |
| title_fullStr | Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes |
| title_full_unstemmed | Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes |
| title_short | Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes |
| title_sort | vibrio natriegens as host for expression of multisubunit membrane protein complexes |
| topic | Vibrio natriegens membrane proteins multisubunit complexes expression Na+-translocating NADH:quinone oxidoreductase multiple resistance and pH related antiporter |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.02537/full |
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