2019
DOI: 10.1105/tpc.18.00854
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A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides

Abstract: The plant endomembrane system facilitates the transport of polysaccharides, associated enzymes, and glycoproteins through its dynamic pathways. Although enzymes involved in cell wall biosynthesis have been identified, little is known about the endomembrane-based transport of glycan components. This is partially attributed to technical challenges in biochemically determining polysaccharide cargo in specific vesicles. Here, we introduce a hybrid approach addressing this limitation. By combining vesicle isolation… Show more

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Cited by 36 publications
(34 citation statements)
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References 118 publications
(166 reference statements)
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“…Given their trans-Golgi association, and the family size, it seems likely that DUF707s make an important contribution to the diversity of terminal substitutions on glycan chains, possibly relating to the cell wall. Nonfucosylated xyloglucan epitopes were not observed in the very latest Golgi cisternae ( Figures 3A to 3C) but have been recorded in post-Golgi compartments and the cell wall (Wilkop et al, 2019). This suggests that their absence from the very late Golgi was not a consequence of further substitution preventing antibody binding.…”
Section: Discussionmentioning
confidence: 80%
See 1 more Smart Citation
“…Given their trans-Golgi association, and the family size, it seems likely that DUF707s make an important contribution to the diversity of terminal substitutions on glycan chains, possibly relating to the cell wall. Nonfucosylated xyloglucan epitopes were not observed in the very latest Golgi cisternae ( Figures 3A to 3C) but have been recorded in post-Golgi compartments and the cell wall (Wilkop et al, 2019). This suggests that their absence from the very late Golgi was not a consequence of further substitution preventing antibody binding.…”
Section: Discussionmentioning
confidence: 80%
“…Following on from the TEM imaging, the FFE fractions (from R1) were analyzed for the same classes of polysaccharide, using carbohydrate antibody arrays immobilized on nitrocellulose membranes, which has been successfully applied to endomembrane enrichments (Okekeogbu et al, 2019) and post-Golgi compartments (Wilkop et al, 2019). Here, we were able to probe an expanded number of polysaccharide epitopes compared with TEM due to the high-throughput nature of the array assays.…”
Section: Cisternal Polysaccharide Distributionmentioning
confidence: 99%
“…It is not clear which vesicle trafficking pathway is disrupted in tno1 mutants, although it appears to involve both SYP41 and SYP61 (Kim and Bassham, 2011). Immunoisolation of vesicles containing SYP61, which may include TGN fragments and TGN-derived vesicles, revealed that they contained secretory cargo including both extracellular proteins and cell wall polysaccharides (Drakakaki et al, 2012;Wilkop et al, 2019). However, there is no evidence to date for a role for TNO1 in secretion, only in transport to the vacuole.…”
Section: Discussionmentioning
confidence: 99%
“…The trans-Golgi network/early endosome (TGN/EE) provides a main trafficking hub for the endomembrane system and comprises a heterogeneous population of vesicles shuttling cargo between Golgi, vacuole, plasma membrane (PM), and late endosomal compartments (Dettmer et al, 2006;Viotti et al, 2010;Wattelet-Boyer et al, 2016;Brillada and Rojas-Pierce, 2017;Rosquete et al, 2018). The plant SYNTAXIN OF PLANTS61 (SYP61) defines a TGN/EE compartment involved in endosomal recycling and transport of extracellular cargo (Robert et al, 2008;Drakakaki et al, 2012;Wilkop et al, 2019) and has been implicated in abiotic stress responses (Zhu et al, 2002).…”
Section: Introductionmentioning
confidence: 99%