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Title: 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 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. Finally, 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.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3]; ORCiD logo [3]; ORCiD logo [3]; ORCiD logo [3]; ORCiD logo [2]; ORCiD logo [4]; ORCiD logo [2]; ORCiD logo [3]
  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)
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States)
Sponsoring Org.:
USDOE Office of Science (SC); National Science Foundation (NSF); USDA; China Scholarship Council
OSTI Identifier:
1612317
Grant/Contract Number:  
SC0015662
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Plant Cell
Additional Journal Information:
Journal Volume: 31; Journal Issue: 3; Journal ID: ISSN 1040-4651
Publisher:
American Society of Plant Biologists
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; biochemistry & molecular biology; plant sciences; cell biology

Citation Formats

Wilkop, Thomas, Pattathil, Sivakumar, Ren, Guangxi, Davis, Destiny J., Bao, Wenlong, Duan, Dechao, Peralta, Angelo G., Domozych, David S., Hahn, Michael G., and Drakakaki, Georgia. A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides. United States: N. p., 2019. Web. doi:10.1105/tpc.18.00854.
Wilkop, Thomas, Pattathil, Sivakumar, Ren, Guangxi, Davis, Destiny J., Bao, Wenlong, Duan, Dechao, Peralta, Angelo G., Domozych, David S., Hahn, Michael G., & Drakakaki, Georgia. A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides. United States. doi:10.1105/tpc.18.00854.
Wilkop, Thomas, Pattathil, Sivakumar, Ren, Guangxi, Davis, Destiny J., Bao, Wenlong, Duan, Dechao, Peralta, Angelo G., Domozych, David S., Hahn, Michael G., and Drakakaki, Georgia. Tue . "A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides". United States. doi:10.1105/tpc.18.00854. https://www.osti.gov/servlets/purl/1612317.
@article{osti_1612317,
title = {A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides},
author = {Wilkop, Thomas and Pattathil, Sivakumar and Ren, Guangxi and Davis, Destiny J. and Bao, Wenlong and Duan, Dechao and Peralta, Angelo G. and Domozych, David S. and Hahn, Michael G. and Drakakaki, Georgia},
abstractNote = {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. Finally, 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.},
doi = {10.1105/tpc.18.00854},
journal = {Plant Cell},
issn = {1040-4651},
number = 3,
volume = 31,
place = {United States},
year = {2019},
month = {4}
}

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