Structural and mechanistic insights into the cleavage of clustered O-glycan patches-containing glycoproteins by mucinases of the human gut

Taleb, Víctor (Universidad de Zaragoza) ; Liao, Qinghua ; Narimatsu, Yoshiki ; García-García, Ana (Universidad de Zaragoza) ; Compañón, Ismael ; Junqueira Borges, Rafael ; González-Ramírez, Andrés Manuel (Universidad de Zaragoza) ; Corzana, Francisco ; Clausen, Henrik ; Rovira, Carme ; Hurtado-Guerrero, Ramon

Resumen: 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.

Idioma: Inglés

DOI: 10.1038/s41467-022-32021-9

Año: 2022

Publicado en: Nature communications 13 (2022), 4324 [15 pp.]
ISSN: 2041-1723

Factor impacto JCR: 16.6 (2022)

Categ. JCR: MULTIDISCIPLINARY SCIENCES rank: 6 / 73 = 0.082 (2022) - Q1 - T1

Factor impacto CITESCORE: 24.9 - Physics and Astronomy (Q1) - General (Q1) - Chemistry (Q1) - Biochemistry, Genetics and Molecular Biology (Q1)

Factor impacto SCIMAGO: 5.116 - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1) - Physics and Astronomy (miscellaneous) (Q1) - Chemistry (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/AEI/BFU2016-75633-P
Financiación: info:eu-repo/grantAgreement/ES/AEI/MDM-2017-0767
Financiación: info:eu-repo/grantAgreement/ES/AEI/PID2019-105451GB-I00
Financiación: info:eu-repo/grantAgreement/ES/AEI/PID2020-118893GB-I00
Financiación: info:eu-repo/grantAgreement/ES/AEI/RTI-2018-099592-B-C21
Financiación: info:eu-repo/grantAgreement/EUR/COST-Action/CA18103-INNOGLY
Financiación: info:eu-repo/grantAgreement/ES/DGA/E34-R17
Financiación: info:eu-repo/grantAgreement/ES/DGA/LMP58-18
Financiación: info:eu-repo/grantAgreement/EC/FP7/283570/EU/Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities/BIOSTRUCT-X
Financiación: info:eu-repo/grantAgreement/EC/H2020/814102/EU/Training interdisciplinary glycoscientists to get a molecular-level grip on glycocodes at the human mucosa–microbiota interface/SWEET CROSSTALK
Financiación: info:eu-repo/grantAgreement/EC/H2020/951231/EU/Activity-Based Profiling of Glycoprocessing Enzymes for Human Health and a Sustainable Society/CARBOCENTRE
Financiación: info:eu-repo/grantAgreement/EC/H2020/956544/EU/Directing the immune response through designed nanomaterials/DIRNANO

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)