Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis
Phosphopantetheinyl transferases (PPTases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPTases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins (CPs), thus orchestrating substrate supply. We describe an efficient...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2016-12-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.12140 |
| _version_ | 1797989108176388096 |
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| author | James Jung Ghader Bashiri Jodie M. Johnston Edward N. Baker |
| author_facet | James Jung Ghader Bashiri Jodie M. Johnston Edward N. Baker |
| author_sort | James Jung |
| collection | DOAJ |
| description | Phosphopantetheinyl transferases (PPTases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPTases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins (CPs), thus orchestrating substrate supply. We describe an efficient mass spectrometry‐based protocol for determining CP specificity for a particular PPTase in organisms possessing several candidate PPTases. We show that the CPs MbtL and PpsC, both involved in synthesis of essential metabolites in Mycobacterium tuberculosis, are exclusively activated by the type 2 PPTase PptT and not the type 1 AcpS. The assay also enables conclusive identification of the reactive serine on each CP. |
| first_indexed | 2024-04-11T08:13:52Z |
| format | Article |
| id | doaj.art-91aca51114004ca599659c1a94461427 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-11T08:13:52Z |
| publishDate | 2016-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-91aca51114004ca599659c1a944614272022-12-22T04:35:14ZengWileyFEBS Open Bio2211-54632016-12-016121220122610.1002/2211-5463.12140Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosisJames Jung0Ghader Bashiri1Jodie M. Johnston2Edward N. Baker3Maurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New ZealandMaurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New ZealandMaurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New ZealandMaurice Wilkins Centre for Molecular Biodiscovery and School of Biological Sciences The University of Auckland New ZealandPhosphopantetheinyl transferases (PPTases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPTases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins (CPs), thus orchestrating substrate supply. We describe an efficient mass spectrometry‐based protocol for determining CP specificity for a particular PPTase in organisms possessing several candidate PPTases. We show that the CPs MbtL and PpsC, both involved in synthesis of essential metabolites in Mycobacterium tuberculosis, are exclusively activated by the type 2 PPTase PptT and not the type 1 AcpS. The assay also enables conclusive identification of the reactive serine on each CP.https://doi.org/10.1002/2211-5463.12140carrier proteinsmass spectrometrymutagenesisMycobacterium tuberculosisphosphopantetheinyl transferases |
| spellingShingle | James Jung Ghader Bashiri Jodie M. Johnston Edward N. Baker Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis FEBS Open Bio carrier proteins mass spectrometry mutagenesis Mycobacterium tuberculosis phosphopantetheinyl transferases |
| title | Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis |
| title_full | Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis |
| title_fullStr | Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis |
| title_full_unstemmed | Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis |
| title_short | Mass spectral determination of phosphopantetheinylation specificity for carrier proteins in Mycobacterium tuberculosis |
| title_sort | mass spectral determination of phosphopantetheinylation specificity for carrier proteins in mycobacterium tuberculosis |
| topic | carrier proteins mass spectrometry mutagenesis Mycobacterium tuberculosis phosphopantetheinyl transferases |
| url | https://doi.org/10.1002/2211-5463.12140 |
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