Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA
The transmembrane domain of the outer membrane protein A (OmpA) from Escherichia coli is an excellent model for structural and folding studies of β-barrel membrane proteins. However, full-length OmpA resists crystallographic efforts, and the link between its function and tertiary structure remains c...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
Cell Press
2014
|
| _version_ | 1826272524384600064 |
|---|---|
| author | Marcoux, J Politis, A Rinehart, D Marshall, D Wallace, M Tamm, L Robinson, C |
| author_facet | Marcoux, J Politis, A Rinehart, D Marshall, D Wallace, M Tamm, L Robinson, C |
| author_sort | Marcoux, J |
| collection | OXFORD |
| description | The transmembrane domain of the outer membrane protein A (OmpA) from Escherichia coli is an excellent model for structural and folding studies of β-barrel membrane proteins. However, full-length OmpA resists crystallographic efforts, and the link between its function and tertiary structure remains controversial. Here we use site-directed mutagenesis and mass spectrometry of different constructs of OmpA, released in the gas phase from detergent micelles, to define the minimal region encompassing the C-terminal dimer interface. Combining knowledge of the location of the dimeric interface with molecular modeling and ion mobility data allows us to propose a low-resolution model for the full-length OmpA dimer. Our model of the dimer is in remarkable agreement with experimental ion mobility data, with none of the unfolding or collapse observed for full-length monomeric OmpA, implying that dimer formation stabilizes the overall structure and prevents collapse of the flexible linker that connects the two domains. © 2014 Elsevier Ltd. |
| first_indexed | 2024-03-06T22:13:55Z |
| format | Journal article |
| id | oxford-uuid:52c53b1f-a1f9-473c-97d1-c6c6c87d1606 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:13:55Z |
| publishDate | 2014 |
| publisher | Cell Press |
| record_format | dspace |
| spelling | oxford-uuid:52c53b1f-a1f9-473c-97d1-c6c6c87d16062022-03-26T16:27:23ZMass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpAJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:52c53b1f-a1f9-473c-97d1-c6c6c87d1606EnglishSymplectic Elements at OxfordCell Press2014Marcoux, JPolitis, ARinehart, DMarshall, DWallace, MTamm, LRobinson, CThe transmembrane domain of the outer membrane protein A (OmpA) from Escherichia coli is an excellent model for structural and folding studies of β-barrel membrane proteins. However, full-length OmpA resists crystallographic efforts, and the link between its function and tertiary structure remains controversial. Here we use site-directed mutagenesis and mass spectrometry of different constructs of OmpA, released in the gas phase from detergent micelles, to define the minimal region encompassing the C-terminal dimer interface. Combining knowledge of the location of the dimeric interface with molecular modeling and ion mobility data allows us to propose a low-resolution model for the full-length OmpA dimer. Our model of the dimer is in remarkable agreement with experimental ion mobility data, with none of the unfolding or collapse observed for full-length monomeric OmpA, implying that dimer formation stabilizes the overall structure and prevents collapse of the flexible linker that connects the two domains. © 2014 Elsevier Ltd. |
| spellingShingle | Marcoux, J Politis, A Rinehart, D Marshall, D Wallace, M Tamm, L Robinson, C Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title | Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title_full | Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title_fullStr | Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title_full_unstemmed | Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title_short | Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA |
| title_sort | mass spectrometry defines the c terminal dimerization domain and enables modeling of the structure of full length ompa |
| work_keys_str_mv | AT marcouxj massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT politisa massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT rinehartd massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT marshalld massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT wallacem massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT tamml massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa AT robinsonc massspectrometrydefinesthecterminaldimerizationdomainandenablesmodelingofthestructureoffulllengthompa |