A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides

Journal Article · Tue Apr 16 00:00:00 EDT 2019 · The Plant Cell

DOI:https://doi.org/10.1105/tpc.18.00854· OSTI ID:1612317

Wilkop, Thomas  ^[1]; Pattathil, Sivakumar  ^[2]; Ren, Guangxi  ^[3]; Davis, Destiny J.  ^[3]; Bao, Wenlong  ^[3]; Duan, Dechao  ^[3]; Peralta, Angelo G.  ^[2]; Domozych, David S.  ^[4]; Hahn, Michael G.  ^[2]; Drakakaki, Georgia  ^[3]

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1. Univ. of California, Davis, CA (United States); Univ. of Kentucky, Lexington, KY (United States)
2. Univ. of Georgia, Athens, GA (United States)
3. Univ. of California, Davis, CA (United States)
4. Skidmore College, Saratoga Springs, NY (United States)

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 with a large-scale carbohydrate antibody arraying technique, we charted an initial large-scale map describing the glycome profile of the SYNTAXIN OF PLANTS61 (SYP61) trans-Golgi network compartment in Arabidopsis (Arabidopsis thaliana). A library of antibodies recognizing specific noncellulosic carbohydrate epitopes allowed us to identify a range of diverse glycans, including pectins, xyloglucans (XyGs), and arabinogalactan proteins in isolated vesicles. Changes in XyG- and pectin-specific epitopes in the cell wall of an Arabidopsis SYP61 mutant corroborate our findings. Our data provide evidence that SYP61 vesicles are involved in the transport and deposition of structural polysaccharides and glycoproteins. Adaptation of our methodology can enable studies characterizing the glycome profiles of various vesicle populations in plant and animal systems and their respective roles in glycan transport defined by subcellular markers, developmental stages, or environmental stimuli.

• Research Organization: Univ. of Georgia, Athens, GA (United States)
• Sponsoring Organization: USDOE Office of Science (SC); National Science Foundation (NSF); USDA; China Scholarship Council
• Grant/Contract Number: SC0015662; IOS 1258135; MCB 1818219; CA-D-PLS-2132-H; DBI-0421683; IOS-0923992
• OSTI ID: 1612317
• Alternate ID(s): OSTI ID: 1964133
• Journal Information: The Plant Cell, Vol. 31, Issue 3; ISSN 1040-4651
• Publisher: American Society of Plant Biologists (ASPB)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
biochemistry & molecular biology
plant sciences
cell biology