Xylosylation of protein N-linked glycans in Chlamydomonas reinhardtii is heterogeneous and mediated by a multigene xylosyltransferase family

Savignien Leprovost (1) , Pierre-Louis Lucas (1) , Chan Philippe (2) , Anne Oltmanns (3) , Corinne Loutelier-Bourhis (4) , Carole Plasson (1) , Carlos Afonso (4) , Patrice Lerouge (1) , Narimane Mati-Baouche (5) , Elodie Mathieu-Rivet (1) , Michael Hippler (3) , Muriel Bardor (1)

1 Glyco-MEV - Laboratoire de Glycobiologie et Matrice Extracellulaire Végétale
2 Plate-forme de Protéomique PISSARO
3 WWU - Westfälische Wilhelms-Universität Münster = University of Münster
4 COBRA - Chimie Organique et Bioorganique : Réactivité et Analyse
5 UGSF - Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576

In eukaryots the protein N-glycosylation processes starts in the endoplasmic reticulum and continues with the maturation steps in the Golgi apparatus (Fig. 1). In C. reinhardtii, the maturation steps results in glycans N-linked to proteins ranging from Man5GlcNAc2 to Man3GlcNAc2 and carrying one or two xylose residues (Fig. 1) (Vanier et al. 2017, Lucas/Dumontier et al. 2018). Nowadays, little information is available regarding the xylosylation in C. reinhardtii. This study aimed at characterizing the molecular actors involved in xylosylation process using complementary analysis, such as Western blot, nanoliquid chromatography coupled to electrospray mass spectrometry (nanoLC-ESI-MS), multistage tandem mass spectrometry (ESI-MSn) and ion mobility spectrometry-tandem mass spectrometry (IMS-MS2), on xylosyltransferase insertional mutants.

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Savignien Leprovost, Pierre-Louis Lucas, Chan Philippe, Anne Oltmanns, Corinne Loutelier-Bourhis, et al.. Xylosylation of protein N-linked glycans in Chlamydomonas reinhardtii is heterogeneous and mediated by a multigene xylosyltransferase family. 19th International Conference on the Cell and Molecular Biology of Chlamydomonas, Aug 2021, Six-Fours-les-Plages, France. ⟨hal-03388367⟩