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Title: Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II

Abstract

Rhamnogalacturonan II (RG-II) is a structurally complex pectic polysaccharide that exists as a borate ester cross-linked dimer in the cell walls of all vascular plants. The glycosyl sequence of RG-II is largely conserved, but there is evidence that galacturonic acid (GalA) methyl etherification and glucuronic acid (GlcA) methyl esterification vary in the A sidechain across plant species. Methyl esterification of the galacturonan backbone has also been reported but not confirmed. Here we describe a new procedure, utilizing aq. sodium borodeuteride (NaBD 4 )-reduced RG-II, to identify the methyl esterification status of backbone GalAs. Our data suggest that up to two different GalAs are esterified in the RG-II backbone. We also adapted a procedure based on methanolysis and NaBD 4 reduction to identify 3-, 4-, and 3,4- O-methyl GalA in RG-II. These data, together with matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (MALDI-TOF) MS analysis of sidechain A generated from selected RG-IIs and their NaBD 4 -reduced counterparts, suggest that methyl etherification of the β-linked GalA and methyl esterification of the GlcA are widespread. Nevertheless, the extent of these modifications varies between plant species. Our analysis of the sidechain B glycoforms in RG-II from different dicots and nonpoalean monocots suggests that this sidechainmore » has a minimum structure of an O-acetylated hexasaccharide (Ara-[MeFuc]-Gal-AceA-Rha-Api-). To complement these studies, we provide further evidence showing that dimer formation and stability in vitro is cation and borate dependent. Taken together, our data further refine the primary sequence and sequence variation of RG-II and provide additional insight into dimer stability and factors controlling dimer self-assembly.« less

Authors:
 [1];  [1];  [1];  [2];  [2];  [1];  [1]
  1. Complex Carbohydrate Research Center, The University of Georgia, Athens, GA, USA
  2. National Renewable Energy Laboratory, Golden, CO, USA
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1631341
Grant/Contract Number:  
SC0008472; SC0015662
Resource Type:
Published Article
Journal Name:
SLAS TECHNOLOGY: Translating Life Sciences Innovation
Additional Journal Information:
Journal Name: SLAS TECHNOLOGY: Translating Life Sciences Innovation Journal Volume: 25 Journal Issue: 4; Journal ID: ISSN 2472-6303
Publisher:
SAGE Publications
Country of Publication:
Country unknown/Code not available
Language:
English

Citation Formats

O’Neill, Malcolm A., Black, Ian, Urbanowicz, Breeanna, Bharadwaj, Vivek, Crowley, Mike, Koj, Sabina, and Peña, Maria J. Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II. Country unknown/Code not available: N. p., 2020. Web. doi:10.1177/2472630320923321.
O’Neill, Malcolm A., Black, Ian, Urbanowicz, Breeanna, Bharadwaj, Vivek, Crowley, Mike, Koj, Sabina, & Peña, Maria J. Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II. Country unknown/Code not available. doi:https://doi.org/10.1177/2472630320923321
O’Neill, Malcolm A., Black, Ian, Urbanowicz, Breeanna, Bharadwaj, Vivek, Crowley, Mike, Koj, Sabina, and Peña, Maria J. Fri . "Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II". Country unknown/Code not available. doi:https://doi.org/10.1177/2472630320923321.
@article{osti_1631341,
title = {Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II},
author = {O’Neill, Malcolm A. and Black, Ian and Urbanowicz, Breeanna and Bharadwaj, Vivek and Crowley, Mike and Koj, Sabina and Peña, Maria J.},
abstractNote = {Rhamnogalacturonan II (RG-II) is a structurally complex pectic polysaccharide that exists as a borate ester cross-linked dimer in the cell walls of all vascular plants. The glycosyl sequence of RG-II is largely conserved, but there is evidence that galacturonic acid (GalA) methyl etherification and glucuronic acid (GlcA) methyl esterification vary in the A sidechain across plant species. Methyl esterification of the galacturonan backbone has also been reported but not confirmed. Here we describe a new procedure, utilizing aq. sodium borodeuteride (NaBD 4 )-reduced RG-II, to identify the methyl esterification status of backbone GalAs. Our data suggest that up to two different GalAs are esterified in the RG-II backbone. We also adapted a procedure based on methanolysis and NaBD 4 reduction to identify 3-, 4-, and 3,4- O-methyl GalA in RG-II. These data, together with matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (MALDI-TOF) MS analysis of sidechain A generated from selected RG-IIs and their NaBD 4 -reduced counterparts, suggest that methyl etherification of the β-linked GalA and methyl esterification of the GlcA are widespread. Nevertheless, the extent of these modifications varies between plant species. Our analysis of the sidechain B glycoforms in RG-II from different dicots and nonpoalean monocots suggests that this sidechain has a minimum structure of an O-acetylated hexasaccharide (Ara-[MeFuc]-Gal-AceA-Rha-Api-). To complement these studies, we provide further evidence showing that dimer formation and stability in vitro is cation and borate dependent. Taken together, our data further refine the primary sequence and sequence variation of RG-II and provide additional insight into dimer stability and factors controlling dimer self-assembly.},
doi = {10.1177/2472630320923321},
journal = {SLAS TECHNOLOGY: Translating Life Sciences Innovation},
number = 4,
volume = 25,
place = {Country unknown/Code not available},
year = {2020},
month = {5}
}

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Works referenced in this record:

Xyloglucan, galactomannan, glucuronoxylan, and rhamnogalacturonan I do not have identical structures in soybean root and root hair cell walls
journal, June 2015


Secondary cell wall characterization in a BY-2 inductive system
journal, July 2013

  • Goué, Nadia; Mortimer, Jenny C.; Nakano, Yoshimi
  • Plant Cell, Tissue and Organ Culture (PCTOC), Vol. 115, Issue 2
  • DOI: 10.1007/s11240-013-0354-7

Silencing of the GDP-d-mannose 3,5-Epimerase Affects the Structure and Cross-linking of the Pectic Polysaccharide Rhamnogalacturonan II and Plant Growth in Tomato
journal, January 2011

  • Voxeur, Aline; Gilbert, Louise; Rihouey, Christophe
  • Journal of Biological Chemistry, Vol. 286, Issue 10
  • DOI: 10.1074/jbc.M110.198614

Symbol Nomenclature for Graphical Representations of Glycans
journal, November 2015

  • Varki, Ajit; Cummings, Richard D.; Aebi, Markus
  • Glycobiology, Vol. 25, Issue 12
  • DOI: 10.1093/glycob/cwv091

Structure of Plant Cell Walls: VIII. A New Pectic Polysaccharide
journal, September 1978

  • Darvill, Alan G.; McNeil, Michael; Albersheim, Peter
  • Plant Physiology, Vol. 62, Issue 3
  • DOI: 10.1104/pp.62.3.418

Characterization of a structurally complex heptasaccharide isolated from the pectic polysaccharide rhamnogalacturonan II
journal, October 1983


The MUR1 gene of Arabidopsis thaliana encodes an isoform of GDP-D-mannose-4,6-dehydratase, catalyzing the first step in the de novo synthesis of GDP-L-fucose
journal, March 1997

  • Bonin, C. P.; Potter, I.; Vanzin, G. F.
  • Proceedings of the National Academy of Sciences, Vol. 94, Issue 5
  • DOI: 10.1073/pnas.94.5.2085

New Insights Into Wall Polysaccharide O-Acetylation
journal, August 2018


Boron uptake by sunflower, squash and cultured tobacco cells
journal, July 1994


Tensile Properties of Arabidopsis Cell Walls Depend on Both a Xyloglucan Cross-Linked Microfibrillar Network and Rhamnogalacturonan II-Borate Complexes
journal, May 2003

  • Ryden, Peter; Sugimoto-Shirasu, Keiko; Smith, Andrew Charles
  • Plant Physiology, Vol. 132, Issue 2
  • DOI: 10.1104/pp.103.021873

Germanium Does Not Substitute for Boron in Cross-Linking of Rhamnogalacturonan II in Pumpkin Cell Walls
journal, November 2002

  • Ishii, Tadashi; Matsunaga, Toshiro; Iwai, Hiroaki
  • Plant Physiology, Vol. 130, Issue 4
  • DOI: 10.1104/pp.009514

A new procedure for the reduction of uronic acid containing polysaccharides
journal, July 1994

  • Fontaine, Thierry; Fournet, Bernard; Karamanos, Yannis
  • Journal of Microbiological Methods, Vol. 20, Issue 2
  • DOI: 10.1016/0167-7012(94)90017-5

The Plant Cell Wall Polysaccharide Rhamnogalacturonan II Self-assembles into a Covalently Cross-linked Dimer
journal, May 1999

  • Ishii, Tadashi; Matsunaga, Toshiro; Pellerin, Patrice
  • Journal of Biological Chemistry, Vol. 274, Issue 19
  • DOI: 10.1074/jbc.274.19.13098

Boron bridging of rhamnogalacturonan-II is promoted in vitro by cationic chaperones, including polyhistidine and wall glycoproteins
journal, August 2015

  • Chormova, Dimitra; Fry, Stephen C.
  • New Phytologist, Vol. 209, Issue 1
  • DOI: 10.1111/nph.13596

RHAMNOGALACTURONAN II: Structure and Function of a Borate Cross-Linked Cell Wall Pectic Polysaccharide
journal, June 2004


Suppression of Arabidopsis GGLT1 affects growth by reducing the L-galactose content and borate cross-linking of rhamnogalacturonan-II
journal, October 2018

  • Sechet, Julien; Htwe, Soe; Urbanowicz, Breeanna
  • The Plant Journal, Vol. 96, Issue 5
  • DOI: 10.1111/tpj.14088

Fruit Softening: Revisiting the Role of Pectin
journal, April 2018


Recovery and fine structure variability of RGII sub-domains in wine (Vitis vinifera Merlot)
journal, June 2014

  • Buffetto, F.; Ropartz, D.; Zhang, X. J.
  • Annals of Botany, Vol. 114, Issue 6
  • DOI: 10.1093/aob/mcu097

Rhamnogalacturonan-II, a Pectic Polysaccharide in the Walls of Growing Plant Cell, Forms a Dimer That Is Covalently Cross-linked by a Borate Ester
journal, September 1996

  • O'Neill, Malcolm A.; Warrenfeltz, Dennis; Kates, Keith
  • Journal of Biological Chemistry, Vol. 271, Issue 37
  • DOI: 10.1074/jbc.271.37.22923

Cellulose-Pectin Spatial Contacts Are Inherent to Never-Dried Arabidopsis Primary Cell Walls: Evidence from Solid-State Nuclear Magnetic Resonance
journal, June 2015

  • Wang, Tuo; Park, Yong Bum; Cosgrove, Daniel J.
  • Plant Physiology, Vol. 168, Issue 3
  • DOI: 10.1104/pp.15.00665

Primary structure of the 2-O-methyl-α-l-fucose-containing side chain of the pectic polysaccharide, rhamnogalacturonan II
journal, February 2003


Structural characterization of red wine rhamnogalacturonan II
journal, September 1996


4-O-methylation of glucuronic acid in Arabidopsis glucuronoxylan is catalyzed by a domain of unknown function family 579 protein
journal, August 2012

  • Urbanowicz, B. R.; Pena, M. J.; Ratnaparkhe, S.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 35
  • DOI: 10.1073/pnas.1208097109

The synthesis of the rhamnogalacturonan II component 3-deoxy-D-manno-2-octulosonic acid (Kdo) is required for pollen tube growth and elongation
journal, July 2008

  • Delmas, F.; Seveno, M.; Northey, J. G. B.
  • Journal of Experimental Botany, Vol. 59, Issue 10
  • DOI: 10.1093/jxb/ern118

Synthesis of borate cross-linked rhamnogalacturonan II
journal, April 2015


Binding of divalent cations to oligomeric fragments of pectin
journal, February 1987


Enhancement of the Quality of MALDI Mass Spectra of Highly Acidic Oligosaccharides by Using a Nafion-Coated Probe
journal, February 2001

  • Jacobs, Anna; Dahlman, Olof
  • Analytical Chemistry, Vol. 73, Issue 3
  • DOI: 10.1021/ac001222i

The Pore Size of Non-Graminaceous Plant Cell Walls Is Rapidly Decreased by Borate Ester Cross-Linking of the Pectic Polysaccharide Rhamnogalacturonan II
journal, November 1999

  • Fleischer, Axel; O'Neill, Malcolm A.; Ehwald, Rudolf
  • Plant Physiology, Vol. 121, Issue 3
  • DOI: 10.1104/pp.121.3.829

Complex pectin metabolism by gut bacteria reveals novel catalytic functions
journal, March 2017

  • Ndeh, Didier; Rogowski, Artur; Cartmell, Alan
  • Nature, Vol. 544, Issue 7648
  • DOI: 10.1038/nature21725

Structural characterisation of the pectic polysaccharide rhamnogalacturonan II using an acidic fingerprinting methodology
journal, August 2009


Determination of the degree of methyl esterification of pectins in small samples by selective reduction of esterified galacturonic acid to galactose
journal, March 1990


Evolving Views of Pectin Biosynthesis
journal, April 2013


A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates
journal, January 1988

  • Domon, Bruno; Costello, Catherine E.
  • Glycoconjugate Journal, Vol. 5, Issue 4
  • DOI: 10.1007/BF01049915

Boron and calcium, essential inorganic constituents of pectic polysaccharides in higher plant cell walls
journal, March 1998

  • Matoh, Toru; Kobayashi, Masaru
  • Journal of Plant Research, Vol. 111, Issue 1
  • DOI: 10.1007/BF02507164

Altered lignification in mur1-1 a mutant deficient in GDP-L-fucose synthesis with reduced RG-II cross linking
journal, September 2017


Depletion of UDP-d-apiose/UDP-d-xylose Synthases Results in Rhamnogalacturonan-II Deficiency, Cell Wall Thickening, and Cell Death in Higher Plants
journal, March 2006

  • Ahn, Joon-Woo; Verma, Rajeev; Kim, Moonil
  • Journal of Biological Chemistry, Vol. 281, Issue 19
  • DOI: 10.1074/jbc.M512403200

Some Aspects of Growth and Metabolism of Paul's Scarlet Rose Cell Suspensions
journal, January 1972

  • Nash, Dudley T.; Davies, M. E.
  • Journal of Experimental Botany, Vol. 23, Issue 1
  • DOI: 10.1093/jxb/23.1.75

Structural characterization of the pectic polysaccharide, rhamnogalacturonan-II
journal, July 1995


Gradients in Wall Mechanics and Polysaccharides along Growing Inflorescence Stems
journal, October 2017

  • Phyo, Pyae; Wang, Tuo; Kiemle, Sarah N.
  • Plant Physiology, Vol. 175, Issue 4
  • DOI: 10.1104/pp.17.01270

Re-constructing our models of cellulose and primary cell wall assembly
journal, December 2014


Boron bridging of rhamnogalacturonan-II, monitored by gel electrophoresis, occurs during polysaccharide synthesis and secretion but not post-secretion
journal, January 2014

  • Chormova, Dimitra; Messenger, David J.; Fry, Stephen C.
  • The Plant Journal, Vol. 77, Issue 4
  • DOI: 10.1111/tpj.12403

Sodium Borohydride, Its Hydrolysis and its Use as a Reducing Agent and in the Generation of Hydrogen 1
journal, January 1953

  • Schlesinger, H. I.; Brown, Herbert C.; Finholt, A. E.
  • Journal of the American Chemical Society, Vol. 75, Issue 1
  • DOI: 10.1021/ja01097a057