Prediction and verification of glycosyltransferase activity by bioinformatics analysis and protein engineering

Dietlind L. Gerloff, Elena I. Ilina, Camille Cialini, Uxue Mata Salcedo, Michel Mittelbronn, Tanja Müller*

*Corresponding author for this work

    Research output: Contribution to journalArticleResearchpeer-review

    1 Citation (Scopus)

    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).1

    Original languageEnglish
    Article number101905
    JournalSTAR Protocols
    Volume4
    Issue number1
    Early online date16 Dec 2022
    DOIs
    Publication statusPublished - 17 Mar 2023

    Keywords

    • Bioinformatics
    • Molecular Biology
    • Protein Biochemistry
    • Protein expression and purification
    • Sequence analysis

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