| Autor: |
Dietlind L. Gerloff, Elena I. Ilina, Camille Cialini, Uxue Mata Salcedo, Michel Mittelbronn, Tanja Müller |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
STAR Protocols, Vol 4, Iss 1, Pp 101905- (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2022.101905 |
| Popis: |
Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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