Efficient TurboID-based proximity labelling method for identifying terminal sialic acid glycosylation in living cells.
Acta Biochim Biophys Sin (Shanghai)
; 54(12): 1841-1853, 2022 Dec 25.
Article
em En
| MEDLINE
| ID: mdl-36789692
ABSTRACT
TurboID, a proximity labelling method based on mutant biotin ligase, is an efficient new technique for recognizing protein-protein interactions and has been successfully applied to living cells. Sialic acid is typically the terminal monosaccharide attached to many glycoproteins and plays many important roles in many biological processes. However, the lack of enrichment methods for terminal sialic acid glycosylation in vivo hinders the identification and analysis of this glycosylation. Here, we introduce TurboID to identify terminal sialic acid glycosylation in living cells. SpCBM, the carbohydrate-binding domain of sialidase from Streptococcus pneumoniae, is fused with TurboID and overexpressed in HeLa cells. After streptavidin-based purification and detection by mass spectrometry, 31 terminal sialic acid N-glycosylated sites and 1359 putative terminal sialic acid glycosylated proteins are identified, many of which are located in the cytoplasm and nucleus.
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Base de dados:
MEDLINE
Assunto principal:
Ácido N-Acetilneuramínico
Limite:
Humans
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article