Structural basis for the synthesis of the core 1 structure by C1GalT1
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Dokumenter
- Fulltext
Forlagets udgivne version, 2,9 MB, PDF-dokument
C1GalT1 is an essential inverting glycosyltransferase responsible for synthesizing the core 1 structure, a common precursor for mucin-type O-glycans found in many glycoproteins. To date, the structure of C1GalT1 and the details of substrate recognition and catalysis remain unknown. Through biophysical and cellular studies, including X-ray crystallography of C1GalT1 complexed to a glycopeptide, we report that C1GalT1 is an obligate GT-A fold dimer that follows a SN2 mechanism. The binding of the glycopeptides to the enzyme is mainly driven by the GalNAc moiety while the peptide sequence provides optimal kinetic and binding parameters. Interestingly, to achieve glycosylation, C1GalT1 recognizes a high-energy conformation of the α-GalNAc-Thr linkage, negligibly populated in solution. By imposing this 3D-arrangement on that fragment, characteristic of α-GalNAc-Ser peptides, C1GalT1 ensures broad glycosylation of both acceptor substrates. These findings illustrate a structural and mechanistic blueprint to explain glycosylation of multiple acceptor substrates, extending the repertoire of mechanisms adopted by glycosyltransferases.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 2398 |
| Tidsskrift | Nature Communications |
| Vol/bind | 13 |
| Udgave nummer | 1 |
| ISSN | 2041-1723 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
We thank the Diamond Light Source (Oxford, UK) synchrotron beamline I24 (experiment number MX20229-11). We thank ARAID, the Agencia Estatal de Investigaci?n (AEI; BFU2016-75633-P and PID2019-105451GB-I00 to R.H-G., and RTI2018-099592-B-C21 to F.C.), Gobierno de Arag?n (E34_R17 and LMP58_18 to R.H-G.) with FEDER (2014?2020) funds for ?Building Europe from Arag?n? for financial support, and the Danish National Research Foundation (DNRF107). F.M., A.S.G. and H.Co. thank to Funda??o para a Ci?ncia e a Tecnologia for funding projects: IF/00780/2015; PTDC/BIA-MIB/31028/2017, UCIBIO project (UIDP/04378/2020 and UIDB/04378/2020) and i4HB project (LA/P/0140/2020). A.S.G. also acknowledges the PhD fellowship (SFRH/BD/140394/2018), and F.M. and H.Co. also thank the CEEC contracts (2020.00233.CEECIND and 2020.03261.CEECIND, respectively). The NMR spectrometers are part of the National NMR Facility supported by FCT-Portugal (ROTEIRO/0031/2013?PINFRA/22161/2016, co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). A.M.G-R. thanks the Spanish Ministry of Science, Innovation and Universities for the FPI fellowship. The research leading to these results has also received funding from the FP7 (2007-2013) under BioStruct-X (grant agreement N?283570 and BIOSTRUCTX_5186).
Funding Information:
We thank the Diamond Light Source (Oxford, UK) synchrotron beamline I24 (experiment number MX20229-11). We thank ARAID, the Agencia Estatal de Investigación (AEI; BFU2016-75633-P and PID2019-105451GB-I00 to R.H-G., and RTI2018-099592-B-C21 to F.C.), Gobierno de Aragón (E34_R17 and LMP58_18 to R.H-G.) with FEDER (2014–2020) funds for “Building Europe from Aragón” for financial support, and the Danish National Research Foundation (DNRF107). F.M., A.S.G. and H.Co. thank to Fundação para a Ciência e a Tecnologia for funding projects: IF/00780/2015; PTDC/BIA-MIB/31028/2017, UCIBIO project (UIDP/04378/2020 and UIDB/04378/2020) and i4HB project (LA/P/0140/2020). A.S.G. also acknowledges the PhD fellowship (SFRH/BD/140394/2018), and F.M. and H.Co. also thank the CEEC contracts (2020.00233.CEECIND and 2020.03261.CEECIND, respectively). The NMR spectrometers are part of the National NMR Facility supported by FCT-Portugal (ROTEIRO/0031/2013–PINFRA/22161/2016, co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). A.M.G-R. thanks the Spanish Ministry of Science, Innovation and Universities for the FPI fellowship. The research leading to these results has also received funding from the FP7 (2007-2013) under BioStruct-X (grant agreement N°283570 and BIOSTRUCTX_5186).
Publisher Copyright:
© 2022, The Author(s).
ID: 306899936