Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism
Abstract Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (S...
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Nature Portfolio
2017-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-04539-2 |
| _version_ | 1828976975615623168 |
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| author | Felix Hirschmann Florian Krause Petra Baruch Igor Chizhov Jonathan Wolf Mueller Dietmar J. Manstein Jutta Papenbrock Roman Fedorov |
| author_facet | Felix Hirschmann Florian Krause Petra Baruch Igor Chizhov Jonathan Wolf Mueller Dietmar J. Manstein Jutta Papenbrock Roman Fedorov |
| author_sort | Felix Hirschmann |
| collection | DOAJ |
| description | Abstract Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (SOT) isoforms were identified. Three of those are involved in glucosinolate (Gl) biosynthesis, glycosylated sulphur-containing aldoximes containing chemically different side chains, whose break-down products are involved in stress response against herbivores, pathogens, and abiotic stress. To explain the differences in substrate specificity of desulpho (ds)-Gl SOTs and to understand the reaction mechanism of plant SOTs, we determined the first high-resolution crystal structure of the plant ds-Gl SOT AtSOT18 in complex with 3′-phosphoadenosine 5′-phosphate (PAP) alone and together with the Gl sinigrin. These new structural insights into the determination of substrate specificity were complemented by mutagenesis studies. The structure of AtSOT18 invigorates the similarity between plant and mammalian sulphotransferases, which illustrates the evolutionary conservation of this multifunctional enzyme family. We identified the essential residues for substrate binding and catalysis and demonstrated that the catalytic mechanism is conserved between human and plant enzymes. Our study indicates that the loop-gating mechanism is likely to be a source of the substrate specificity in plants. |
| first_indexed | 2024-12-14T14:49:26Z |
| format | Article |
| id | doaj.art-7759b0cf7dcd4567b7acaaa342f0ca1a |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-14T14:49:26Z |
| publishDate | 2017-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-7759b0cf7dcd4567b7acaaa342f0ca1a2022-12-21T22:57:11ZengNature PortfolioScientific Reports2045-23222017-06-017111210.1038/s41598-017-04539-2Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanismFelix Hirschmann0Florian Krause1Petra Baruch2Igor Chizhov3Jonathan Wolf Mueller4Dietmar J. Manstein5Jutta Papenbrock6Roman Fedorov7Institute of Botany, Leibniz University HannoverInstitute of Botany, Leibniz University HannoverInstitute for Biophysical Chemistry, Hannover Medical SchoolInstitute for Biophysical Chemistry, Hannover Medical SchoolInstitute of Metabolism and Systems Research (IMSR), University of BirminghamInstitute for Biophysical Chemistry, Hannover Medical SchoolInstitute of Botany, Leibniz University HannoverInstitute for Biophysical Chemistry, Hannover Medical SchoolAbstract Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (SOT) isoforms were identified. Three of those are involved in glucosinolate (Gl) biosynthesis, glycosylated sulphur-containing aldoximes containing chemically different side chains, whose break-down products are involved in stress response against herbivores, pathogens, and abiotic stress. To explain the differences in substrate specificity of desulpho (ds)-Gl SOTs and to understand the reaction mechanism of plant SOTs, we determined the first high-resolution crystal structure of the plant ds-Gl SOT AtSOT18 in complex with 3′-phosphoadenosine 5′-phosphate (PAP) alone and together with the Gl sinigrin. These new structural insights into the determination of substrate specificity were complemented by mutagenesis studies. The structure of AtSOT18 invigorates the similarity between plant and mammalian sulphotransferases, which illustrates the evolutionary conservation of this multifunctional enzyme family. We identified the essential residues for substrate binding and catalysis and demonstrated that the catalytic mechanism is conserved between human and plant enzymes. Our study indicates that the loop-gating mechanism is likely to be a source of the substrate specificity in plants.https://doi.org/10.1038/s41598-017-04539-2 |
| spellingShingle | Felix Hirschmann Florian Krause Petra Baruch Igor Chizhov Jonathan Wolf Mueller Dietmar J. Manstein Jutta Papenbrock Roman Fedorov Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism Scientific Reports |
| title | Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| title_full | Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| title_fullStr | Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| title_full_unstemmed | Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| title_short | Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| title_sort | structural and biochemical studies of sulphotransferase 18 from arabidopsis thaliana explain its substrate specificity and reaction mechanism |
| url | https://doi.org/10.1038/s41598-017-04539-2 |
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