Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB
Abstract Non-ribosomal peptide synthetases (NRPS) are multi-modular/domain enzymes that catalyze the synthesis of bioactive peptides. A crucial step in the process is peptide elongation accomplished by the condensation (C) domain with the aid of a peptidyl carrier or thiolation (T) domain. Here, we...
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| Format: | Article |
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Nature Portfolio
2022-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-09188-8 |
| _version_ | 1818774183204618240 |
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| author | Melissa-Jane Chu Yuan Kee Sakshibeedu R. Bharath Sheena Wee Matthew W. Bowler Jayantha Gunaratne Shenquan Pan Lianhui Zhang Haiwei Song |
| author_facet | Melissa-Jane Chu Yuan Kee Sakshibeedu R. Bharath Sheena Wee Matthew W. Bowler Jayantha Gunaratne Shenquan Pan Lianhui Zhang Haiwei Song |
| author_sort | Melissa-Jane Chu Yuan Kee |
| collection | DOAJ |
| description | Abstract Non-ribosomal peptide synthetases (NRPS) are multi-modular/domain enzymes that catalyze the synthesis of bioactive peptides. A crucial step in the process is peptide elongation accomplished by the condensation (C) domain with the aid of a peptidyl carrier or thiolation (T) domain. Here, we examined condensation reaction carried out by NRPS AmbB involved in biosynthesis of l-2-amino-4-methoxy-trans-3-butenoic acid (AMB) in P. aeruginosa. We determined crystal structures of the truncated T–C bidomain of AmbB in three forms, the apo enzyme with disordered T domain, the holo form with serine linked phosphopantetheine (Ppant) and a holo form with substrate (l-alanine) loaded onto Ppant. The two holo forms feature the T domain in a substrate-donation conformation. Mutagenesis combined with functional assays identified residues essential for the attachment of Ppant, anchoring the Ppant-l-Ala in the donor catalytic channel and the role of the conserved His953 in condensation activity. Altogether, these results provide structural insights into the condensation reaction at the donor site with a substrate-bound C domain of AmbB and lay the foundation for understanding the molecular mechanism of condensation which is crucial for AMB synthesis. |
| first_indexed | 2024-12-18T10:37:05Z |
| format | Article |
| id | doaj.art-e3398d8665514b78aa5086f2006efa1e |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-18T10:37:05Z |
| publishDate | 2022-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
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| spelling | doaj.art-e3398d8665514b78aa5086f2006efa1e2022-12-21T21:10:44ZengNature PortfolioScientific Reports2045-23222022-03-0112111310.1038/s41598-022-09188-8Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbBMelissa-Jane Chu Yuan Kee0Sakshibeedu R. Bharath1Sheena Wee2Matthew W. Bowler3Jayantha Gunaratne4Shenquan Pan5Lianhui Zhang6Haiwei Song7Institute of Molecular and Cell BiologyInstitute of Molecular and Cell BiologyInstitute of Molecular and Cell BiologyEuropean Molecular Biology LaboratoryInstitute of Molecular and Cell BiologyDepartment of Biological Sciences, National University of SingaporeSouth China Agricultural UniversityInstitute of Molecular and Cell BiologyAbstract Non-ribosomal peptide synthetases (NRPS) are multi-modular/domain enzymes that catalyze the synthesis of bioactive peptides. A crucial step in the process is peptide elongation accomplished by the condensation (C) domain with the aid of a peptidyl carrier or thiolation (T) domain. Here, we examined condensation reaction carried out by NRPS AmbB involved in biosynthesis of l-2-amino-4-methoxy-trans-3-butenoic acid (AMB) in P. aeruginosa. We determined crystal structures of the truncated T–C bidomain of AmbB in three forms, the apo enzyme with disordered T domain, the holo form with serine linked phosphopantetheine (Ppant) and a holo form with substrate (l-alanine) loaded onto Ppant. The two holo forms feature the T domain in a substrate-donation conformation. Mutagenesis combined with functional assays identified residues essential for the attachment of Ppant, anchoring the Ppant-l-Ala in the donor catalytic channel and the role of the conserved His953 in condensation activity. Altogether, these results provide structural insights into the condensation reaction at the donor site with a substrate-bound C domain of AmbB and lay the foundation for understanding the molecular mechanism of condensation which is crucial for AMB synthesis.https://doi.org/10.1038/s41598-022-09188-8 |
| spellingShingle | Melissa-Jane Chu Yuan Kee Sakshibeedu R. Bharath Sheena Wee Matthew W. Bowler Jayantha Gunaratne Shenquan Pan Lianhui Zhang Haiwei Song Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB Scientific Reports |
| title | Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB |
| title_full | Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB |
| title_fullStr | Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB |
| title_full_unstemmed | Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB |
| title_short | Structural insights into the substrate-bound condensation domains of non-ribosomal peptide synthetase AmbB |
| title_sort | structural insights into the substrate bound condensation domains of non ribosomal peptide synthetase ambb |
| url | https://doi.org/10.1038/s41598-022-09188-8 |
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