{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["4(1)"],"submitter":["Gerloff DL"],"funding":["Fonds National de la Recherche Luxembourg"],"pubmed_abstract":["A significant number of proteins are annotated as functionally uncharacterized proteins. Within this protocol, we describe how to use protein family multiple sequence alignments and structural bioinformatics resources to design loss-of-function mutations of previously uncharacterized proteins within the glycosyltransferase family. We detail approaches to determine target protein active sites using three-dimensional modeling. We generate active site mutants and quantify any changes in enzymatic function by a glycosyltransferase assay. With modifications, this protocol could be applied to other metal-dependent enzymes. For complete details on the use and execution of this protocol, please refer to Ilina et al. (2022).<sup>1</sup>."],"journal":["STAR protocols"],"pagination":["101905"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9792956"],"repository":["biostudies-literature"],"pubmed_title":["Prediction and verification of glycosyltransferase activity by bioinformatics analysis and protein engineering."],"pmcid":["PMC9792956"],"pubmed_authors":["Mata Salcedo U","Cialini C","Ilina EI","Muller T","Gerloff DL","Mittelbronn M"],"additional_accession":[]},"is_claimable":false,"name":"Prediction and verification of glycosyltransferase activity by bioinformatics analysis and protein engineering.","description":"A significant number of proteins are annotated as functionally uncharacterized proteins. Within this protocol, we describe how to use protein family multiple sequence alignments and structural bioinformatics resources to design loss-of-function mutations of previously uncharacterized proteins within the glycosyltransferase family. We detail approaches to determine target protein active sites using three-dimensional modeling. We generate active site mutants and quantify any changes in enzymatic function by a glycosyltransferase assay. With modifications, this protocol could be applied to other metal-dependent enzymes. For complete details on the use and execution of this protocol, please refer to Ilina et al. (2022).<sup>1</sup>.","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Mar","modification":"2025-04-05T14:54:59.81Z","creation":"2024-11-13T09:37:14.349Z"},"accession":"S-EPMC9792956","cross_references":{"pubmed":["36528856"],"doi":["10.1016/j.xpro.2022.101905"]}}