| Title: |
A Rapid and Sensitive MicroPlate Assay (MPSA) Using an Alkyne‐Modified CMP‐Sialic Acid Donor to Evaluate Human Sialyltransferase Specificity |
| Authors: |
Clairene Filipe, Kiamungongo; Dangudubiyyam, Sushmaa; Lion, Cedric; Decloquement, Mathieu; Teppa, Roxana; Biot, Christophe; Harduin-Lepers, Anne |
| Contributors: |
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF); Université de Lille-Centre National de la Recherche Scientifique (CNRS); ANR-21-CE44-0032,PsaMAr,Des polysialyltransférases d'origine marine pour la synthèse de bioconjugués immunomodulateurs(2021) |
| Source: |
ISSN: 1439-4227. |
| Publisher Information: |
CCSD; Wiley-VCH Verlag |
| Publication Year: |
2024 |
| Collection: |
LillOA (HAL Lille Open Archive, Université de Lille) |
| Subject Terms: |
Substrate specificity; Sialyltransferase; Glycobiology; Enzyme assay; Bioorthognal chemistry; [SDV]Life Sciences [q-bio]; [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry |
| Description: |
International audience ; Human sialyltransferases primarily utilize CMP‐Sias, especially transferring Neu5Ac from CMP‐Neu5Ac to various acceptors. Advances in chemical biology have led to the synthesis of novel CMP‐Sia donors suitable for bioorthogonal reactions in cell‐based assays. However, the compatibility of these donors with all human enzymes remains uncertain. We synthesized a non‐natural CMP‐Sia donor with an alkyne modification on the N‐acyl group of Neu5Ac, which was effectively used by human ST6Gal I and ST3Gal I. A sensitive MicroPlate Sialyltransferase Assay (MPSA) was developed and expanded to a panel of 13 human STs acting on glycoproteins. All assayed enzymes tolerated CMP‐SiaNAl, allowing for the determination of kinetic parameters and turnover numbers. This study enhances the biochemical characterization of human sialyltransferases and opens new avenues for developing sialyltransferase inhibitors. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1002/cbic.202400539 |
| Availability: |
https://lilloa.hal.science/hal-04835900; https://lilloa.hal.science/hal-04835900v1/document; https://lilloa.hal.science/hal-04835900v1/file/P24.19%20FILIPE%20Harduin-Lepers%20ChemBioChem%20-%202024%20-%20MPSA%20%20Using%20an%20Alkyne%E2%80%90Modified%20CMP%E2%80%90Sialic.pdf; https://doi.org/10.1002/cbic.202400539 |
| Rights: |
https://creativecommons.org/licenses/by/4.0/ ; info:eu-repo/semantics/OpenAccess |
| Accession Number: |
edsbas.9D02405E |
| Database: |
BASE |