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. 2021 Mar 17;12:638454. doi: 10.3389/fpls.2021.638454

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Copyright © 2021 Alvisi, van Noort, Dwiani, Geschiere, Sukarta, Varossieau, Nguyen, Strasser, Hokke, Schots and Wilbers.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Cleavage of engineered LDN motifs by N. benthamiana HEXOs. Kappa-5 was co-expressed with p19, CeGalNAcT and different HEXO genes in wild-type plants. After isolation of the apoplast fluid (at 6 dpi) the presence of the LDN motif on kappa-5 N-glycans was analyzed. (A) Soybean agglutinin (SBA) binding assay for detection of terminal GalNAc. Lectin binding is measured as absorbance at 450 nm. (B–D) Kappa-5 was purified from the apoplast fluid and the N-glycan composition was analyzed by MALDI-TOF-MS. MS profiles are given for N-glycans of kappa-5 upon co-expression of GalNAcT only (B), or combined with co-expression of HEXO2B (C), or HEXO3A (D). When a MS peak represents multiple N-glycan structures of identical mass, the possible positions of these sugar residues on the N-glycan are indicated between brackets. m/z refers to mass-to-charge ratio.