{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Sato M"],"funding":["NIGMS NIH HHS"],"pagination":["223-232"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9302879"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["4(3)"],"pubmed_abstract":["We have previously reported the identification of CghA, a proposed Diels-Alderase responsible for the formation of the bicyclic octalin core of the fungal secondary metabolite Sch210972. Here we show the crystal structure of the CghA-product complex at a resolution of 2.0 Å. Our result provides the second structural determination of eukaryotic Diels-Alderases and adds yet another fold to the family of proteins reported to catalyse [4 + 2] cycloaddition reactions. Site-directed mutagenesis-coupled kinetic characterization and computational analyses allowed us to identify key catalytic residues and propose a possible catalytic mechanism. Most interestingly, we were able to rationally engineer CghA such that the mutant was able to catalyse preferentially the formation of the energetically disfavoured <i>exo</i> adduct. This work expands our knowledge and understanding of the emerging and potentially widespread class of natural enzymes capable of catalysing stereoselective Diels-Alder reactions and paves the way towards developing enzymes potentially useful in various bio/synthetic applications."],"journal":["Nature catalysis"],"pubmed_title":["Catalytic mechanism and <i>endo</i>-to-<i>exo</i> selectivity reversion of an octalin-forming natural Diels-Alderase."],"pmcid":["PMC9302879"],"funding_grant_id":["R01 GM124480"],"pubmed_authors":["Tsunematsu Y","Maeda N","Sato M","Yokoyama M","Hara K","Houk KN","Watanabe K","Narita K","Kishimoto S","Tang Y","Jamieson CS","Hashimoto H"],"additional_accession":[]},"is_claimable":false,"name":"Catalytic mechanism and <i>endo</i>-to-<i>exo</i> selectivity reversion of an octalin-forming natural Diels-Alderase.","description":"We have previously reported the identification of CghA, a proposed Diels-Alderase responsible for the formation of the bicyclic octalin core of the fungal secondary metabolite Sch210972. Here we show the crystal structure of the CghA-product complex at a resolution of 2.0 Å. Our result provides the second structural determination of eukaryotic Diels-Alderases and adds yet another fold to the family of proteins reported to catalyse [4 + 2] cycloaddition reactions. Site-directed mutagenesis-coupled kinetic characterization and computational analyses allowed us to identify key catalytic residues and propose a possible catalytic mechanism. Most interestingly, we were able to rationally engineer CghA such that the mutant was able to catalyse preferentially the formation of the energetically disfavoured <i>exo</i> adduct. This work expands our knowledge and understanding of the emerging and potentially widespread class of natural enzymes capable of catalysing stereoselective Diels-Alder reactions and paves the way towards developing enzymes potentially useful in various bio/synthetic applications.","dates":{"release":"2021-01-01T00:00:00Z","publication":"2021 Mar","modification":"2025-04-25T19:45:12.517Z","creation":"2025-04-06T08:09:50.147Z"},"accession":"S-EPMC9302879","cross_references":{"pubmed":["35873532"],"doi":["10.1038/s41929-021-00577-2"]}}