Structural and mechanistic insights into the cleavage of clustered O-glycan patches-containing glycoproteins by mucinases of the human gut
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Mucinases of human gut bacteria cleave peptide bonds in mucins strictly depending on the presence of neighboring O-glycans. The Akkermansia muciniphila AM0627 mucinase cleaves specifically in between contiguous (bis) O-glycans of defined truncated structures, suggesting that this enzyme may recognize clustered O-glycan patches. Here, we report the structure and molecular mechanism of AM0627 in complex with a glycopeptide containing a bis-T (Galβ1-3GalNAcα1-O-Ser/Thr) O-glycan, revealing that AM0627 recognizes both the sugar moieties and the peptide sequence. AM0627 exhibits preference for bis-T over bis-Tn (GalNAcα1-O-Ser/Thr) O-glycopeptide substrates, with the first GalNAc residue being essential for cleavage. AM0627 follows a mechanism relying on a nucleophilic water molecule and a catalytic base Glu residue. Structural comparison among mucinases identifies a conserved Tyr engaged in sugar-π interactions in both AM0627 and the Bacteroides thetaiotaomicron BT4244 mucinase as responsible for the common activity of these two mucinases with bis-T/Tn substrates. Our work illustrates how mucinases through tremendous flexibility adapt to the diversity in distribution and patterns of O-glycans on mucins.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 4324 |
| Tidsskrift | Nature Communications |
| Vol/bind | 13 |
| Udgave nummer | 1 |
| ISSN | 2041-1723 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
We thank the ALBA (Barcelona, Spain) synchrotron beamline XALOC. We thank ARAID, the Agencia Estatal de Investigación (AEI, BFU2016-75633-P and PID2019-105451GB-I00 to R.H.-G., RTI2018-099592-B-C21 to F.C., PID2020-118893GB-I00 to C.R.), the Fondo Europeo de Desarrollo Regional (FEDER), the Spanish Structures of Excellence María de Maeztu (MDM-2017-0767 to C.R.), 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, the Lundbeck Foundation, the Novo Nordisk Foundation, and the Danish National Research Foundation (DNRF107) to H.C.), the COST Action CA18103 INNOGLY: Innovation with Glycans new frontiers from synthesis to new biological targets, the European Research Council (ERC-2020-SyG-951231 “CARBOCENTRE” to C.R.) and the Marie Skłodowska-Curie Innovative Training Networks (H2020-MSCA-ITN-2018-814102 “Sweet Crosstalk” to C.R.). V.T. thanks the Spanish Ministry of Science, Innovation and Universities for the FPI fellowship. Q.L. thanks AGAUR for the Beatriu de Pinos postdoctoral fellowship (No. 2019 BP 00129). RJB thanks to FAPESP (2016/24191-8). Y.N. thanks the Mizutani Foundation for Glycoscience (grant 210086). The research leading to these results has also received funding from the FP7 (2007-2013) under BioStruct-X (grant agreement No. 283570 and BIOSTRUCTX_5186). F.C. thanks the Mizutani Foundation for Glycoscience (grant 220115) and the EU (Marie-Sklodowska Curie ITN, DIRNANO, grant agreement No. 956544).
Funding Information:
We thank the ALBA (Barcelona, Spain) synchrotron beamline XALOC. We thank ARAID, the Agencia Estatal de Investigación (AEI, BFU2016-75633-P and PID2019-105451GB-I00 to R.H.-G., RTI2018-099592-B-C21 to F.C., PID2020-118893GB-I00 to C.R.), the Fondo Europeo de Desarrollo Regional (FEDER), the Spanish Structures of Excellence María de Maeztu (MDM-2017-0767 to C.R.), 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, the Lundbeck Foundation, the Novo Nordisk Foundation, and the Danish National Research Foundation (DNRF107) to H.C.), the COST Action CA18103 INNOGLY: Innovation with Glycans new frontiers from synthesis to new biological targets, the European Research Council (ERC-2020-SyG-951231 “CARBOCENTRE” to C.R.) and the Marie Skłodowska-Curie Innovative Training Networks (H2020-MSCA-ITN-2018-814102 “Sweet Crosstalk” to C.R.). V.T. thanks the Spanish Ministry of Science, Innovation and Universities for the FPI fellowship. Q.L. thanks AGAUR for the Beatriu de Pinos postdoctoral fellowship (No. 2019 BP 00129). RJB thanks to FAPESP (2016/24191-8). Y.N. thanks the Mizutani Foundation for Glycoscience (grant 210086). The research leading to these results has also received funding from the FP7 (2007-2013) under BioStruct-X (grant agreement No. 283570 and BIOSTRUCTX_5186). F.C. thanks the Mizutani Foundation for Glycoscience (grant 220115) and the EU (Marie-Sklodowska Curie ITN, DIRNANO, grant agreement No. 956544).
Publisher Copyright:
© 2022, The Author(s).
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