19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase
Resistance to β-lactam antibiotics mediated by metallo-β-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19F-NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM-1) from β-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, w...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Wiley
2017
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| _version_ | 1826307681674067968 |
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| author | Abboud, M Hinchliffe, P Brem, J Macsics, R Pfeffer, I Makena, A Umland, K Rydzik, A Li, G Spencer, J Claridge, T Schofield, C |
| author_facet | Abboud, M Hinchliffe, P Brem, J Macsics, R Pfeffer, I Makena, A Umland, K Rydzik, A Li, G Spencer, J Claridge, T Schofield, C |
| author_sort | Abboud, M |
| collection | OXFORD |
| description | Resistance to β-lactam antibiotics mediated by metallo-β-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19F-NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM-1) from β-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di-ZnII active site, were selectively 19F-labeled using 3-bromo-1,1,1-trifluoroacetone. The PrOF NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed β-lactam products to SPM-1. These results have implications for the mechanisms and inhibition of MBLs by β-lactams and non-β-lactams and illustrate the utility of PrOF NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers. |
| first_indexed | 2024-03-07T07:08:23Z |
| format | Journal article |
| id | oxford-uuid:d218ee2e-bbbd-40ce-8bd9-c9bd0df99575 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:08:23Z |
| publishDate | 2017 |
| publisher | Wiley |
| record_format | dspace |
| spelling | oxford-uuid:d218ee2e-bbbd-40ce-8bd9-c9bd0df995752022-05-17T19:27:43Z19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamaseJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d218ee2e-bbbd-40ce-8bd9-c9bd0df99575EnglishSymplectic Elements at OxfordWiley2017Abboud, MHinchliffe, PBrem, JMacsics, RPfeffer, IMakena, AUmland, KRydzik, ALi, GSpencer, JClaridge, TSchofield, CResistance to β-lactam antibiotics mediated by metallo-β-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19F-NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM-1) from β-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di-ZnII active site, were selectively 19F-labeled using 3-bromo-1,1,1-trifluoroacetone. The PrOF NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed β-lactam products to SPM-1. These results have implications for the mechanisms and inhibition of MBLs by β-lactams and non-β-lactams and illustrate the utility of PrOF NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers. |
| spellingShingle | Abboud, M Hinchliffe, P Brem, J Macsics, R Pfeffer, I Makena, A Umland, K Rydzik, A Li, G Spencer, J Claridge, T Schofield, C 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title | 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title_full | 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title_fullStr | 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title_full_unstemmed | 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title_short | 19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paolo metallo-β-lactamase |
| title_sort | 19f nmr reveals the role of mobile loops in product and inhibitor binding by the sao paolo metallo β lactamase |
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