000123962 001__ 123962
000123962 005__ 20230914083757.0
000123962 0247_ $$2doi$$a10.1021/jacsau.1c00529
000123962 0248_ $$2sideral$$a132679
000123962 037__ $$aART-2022-132679
000123962 041__ $$aeng
000123962 100__ $$aCoelho, Helena
000123962 245__ $$aAtomic and specificity details of Mucin 1 O-Glycosylation process by Multiple Polypeptide GalNAc-Transferase Isoforms unveiled by NMR and Molecular Modeling
000123962 260__ $$c2022
000123962 5060_ $$aAccess copy available to the general public$$fUnrestricted
000123962 5203_ $$aThe large family of polypeptide GalNAc-transferases (GalNAc-Ts) controls with precision how GalNAc O-glycans are added in the tandem repeat regions of mucins (e.g., MUC1). However, the structural features behind the creation of well-defined and clustered patterns of O-glycans in mucins are poorly understood. In this context, herein, we disclose the full process of MUC1 O-glycosylation by GalNAc-T2/T3/T4 isoforms by NMR spectroscopy assisted by molecular modeling protocols. By using MUC1, with four tandem repeat domains as a substrate, we confirmed the glycosylation preferences of different GalNAc-Ts isoforms and highlighted the importance of the lectin domain in the glycosylation site selection after the addition of the first GalNAc residue. In a glycosylated substrate, with yet multiple acceptor sites, the lectin domain contributes to orientate acceptor sites to the catalytic domain. Our experiments suggest that during this process, neighboring tandem repeats are critical for further glycosylation of acceptor sites by GalNAc-T2/T4 in a lectin-assisted manner. Our studies also show local conformational changes in the peptide backbone during incorporation of GalNAc residues, which might explain GalNAc-T2/T3/T4 fine specificities toward the MUC1 substrate. Interestingly, we postulate that a specific salt-bridge and the inverse γ-turn conformation of the PDTRP sequence in MUC1 are the main structural motifs behind the GalNAc-T4 specificity toward this region. In addition, in-cell analysis shows that the GalNAc-T4 isoform is the only isoform glycosylating the Thr of the immunogenic epitope PDTRP in vivo, which highlights the relevance of GalNAc-T4 in the glycosylation of this epitope. Finally, the NMR methodology established herein can be extended to other glycosyltransferases, such as C1GalT1 and ST6GalNAc-I, to determine the specificity toward complex mucin acceptor substrates.
000123962 536__ $$9info:eu-repo/grantAgreement/ES/AEI/RTI-2018-099592-B-C21$$9info:eu-repo/grantAgreement/ES/DGA/E34-R17$$9info:eu-repo/grantAgreement/ES/DGA/LMP58-18$$9info:eu-repo/grantAgreement/EC/H2020/956544/EU/Directing the immune response through designed nanomaterials/DIRNANO$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 956544-DIRNANO$$9info:eu-repo/grantAgreement/ES/MICINN/RTI2018-094751-B-C21
000123962 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000123962 594__ $$a5.3$$b2022
000123962 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000123962 700__ $$0(orcid)0000-0002-1924-334X$$aRivas, Matilde de las
000123962 700__ $$aGrosso, Ana S.
000123962 700__ $$aDiniz, Ana
000123962 700__ $$aSoares, Cátia O.
000123962 700__ $$aFrancisco, Rodrigo A.
000123962 700__ $$aDias, Jorge S.
000123962 700__ $$aCompañon, Ismael
000123962 700__ $$aSun, Lingbo
000123962 700__ $$aNarimatsu, Yoshiki
000123962 700__ $$aVakhrushev, Sergey Y.
000123962 700__ $$aClausen, Henrik
000123962 700__ $$aCabrita, Eurico J.
000123962 700__ $$aJiménez-Barbero, Jesús
000123962 700__ $$aCorzana, Francisco
000123962 700__ $$0(orcid)0000-0002-3122-9401$$aHurtado-Guerrero, Ramon
000123962 700__ $$aMarcelo, Filipa
000123962 773__ $$g2, 3 (2022), 631-645$$tJACS Au$$x2691-3704
000123962 8564_ $$s7457632$$uhttps://zaguan.unizar.es/record/123962/files/texto_completo.pdf$$yVersión publicada
000123962 8564_ $$s3202767$$uhttps://zaguan.unizar.es/record/123962/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000123962 909CO $$ooai:zaguan.unizar.es:123962$$particulos$$pdriver
000123962 951__ $$a2023-09-13-15:17:44
000123962 980__ $$aARTICLE