Arabinogalactan protein and wall-associated kinase in a plasmalemmal reticulum with specialized vertices

PROTOPLASMA ◽  
2000 ◽  
Vol 212 (1-2) ◽  
pp. 115-134 ◽  
Author(s):  
J. Scott Gens ◽  
Masaaki Fujiki ◽  
Barbara G. Pickard
Keyword(s):  
Arabinogalactan Protein ◽  
Plasmalemmal Reticulum

Ca2+ pulsation in BY-2 cells and evidence for control of mechanosensory Ca2+-selective channels by the plasmalemmal reticulum

2005 ◽  
Vol 32 (10) ◽  
pp. 863 ◽  
Author(s):  
Barbara G. Pickard ◽  
Masaaki Fujiki
Keyword(s):  
Cell Cycle ◽  
Signal Transduction ◽  
Arabinogalactan Protein ◽  
Cation Channels ◽  
Carrier Solution ◽  
Cytoskeletal Structure ◽  
Yariv Reagent ◽  
Yariv Phenylglycoside ◽  
Plasmalemmal Reticulum ◽  
Oscillation Rate

A previously unknown cytoskeletal structure, now named the plasmalemmal reticulum (Gens et al. 2000, Protoplasma 212, 115–134), was found in cultured BY-2 tobacco cells during a search for a force-focusing mechanism that might enhance signal transduction by the cells’ mechanosensory Ca2+-selective cation channels (MCaCs). This polyhedral structure, which links cell wall, plasma membrane, and internal cytoplasm, prominently contains arabinogalactan protein (AGP). To check for reticulum-promoted Ca2+ elevation, the AGP-binding reagent (β-d-glucosyl)3 Yariv phenylglycoside has been applied to BY-2 cells expressing a free cameleon Ca2+ reporter. Ca2+ elevation was substantial and prolonged. Moreover it occurred in the nucleus as well as the cytoplasm. Cells treated with non-binding mannosyl Yariv reagent could not be discriminated from untreated controls or those treated with carrier solution alone. Supply of the MCaC inhibiter Gd3+ just before treatment with Yariv reagent prevented Ca2+ rise. These data strongly support the hypothesis that the plasmalemmal reticulum controls MCaC activity. The massive inward spread of Ca2+ suggested that entry of the ion through the channels initiated a wave of release from the ER, and YCX in the ER showed Ca2+ levels consistent with this premise. Cytosolic and nuclear Ca2+ often pulsed in control cells in near synchrony and at rates ranging from zero to five cycles per ∼20-min recording. (Pulsation was over-ridden by the applied amounts of glucosyl Yariv compound.) Suggestively but very crudely, oscillation rate was assessed as possibly correlating with stage of cell cycle. Because cell Ca2+ was lowered and pulsation was eliminated by Gd3+, MCaCs appear to participate in these endogenous fluctuations. The extent to which pulsing plays regulatory roles in relatively undifferentiated types of cells should be evaluated.

Characterisation of extracellular polysaccharides from suspension cultures of members of the Poaceae

2020 ◽  
Author(s):  
Ian Sims ◽  
K Middleton ◽  
AG Lane ◽  
AJ Cairns ◽  
A Bacic
Keyword(s):  
Cultured Cells ◽  
Oryza Sativa L ◽  
Suspension Cultures ◽  
Exchange Chromatography ◽  
Phleum Pratense ◽  
Arabinogalactan Protein ◽  
Extracellular Polysaccharides ◽  
Type Ii ◽  
Hordeum Vulgare L ◽  
Suspension Cultured Cells

Microscopic examination of suspension-cultured cells of Phleum pratense L., Panicum miliaceum L., Phalaris aquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Characterisation of extracellular polysaccharides from suspension cultures of members of the Poaceae

2020 ◽  
Author(s):  
Ian Sims ◽  
K Middleton ◽  
AG Lane ◽  
AJ Cairns ◽  
A Bacic
Keyword(s):  
Cultured Cells ◽  
Oryza Sativa L ◽  
Suspension Cultures ◽  
Exchange Chromatography ◽  
Phleum Pratense ◽  
Arabinogalactan Protein ◽  
Extracellular Polysaccharides ◽  
Type Ii ◽  
Hordeum Vulgare L ◽  
Suspension Cultured Cells

Microscopic examination of suspension-cultured cells of Phleum pratense L., Panicum miliaceum L., Phalaris aquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Structure of the exudate gum from Meryta sinclairii

2020 ◽  
Author(s):  
Ian Sims ◽  
Richard Furneaux
Keyword(s):  
Average Molecular Weight ◽  
Gum Arabic ◽  
Arabinogalactan Protein ◽  
Weight Average Molecular Weight ◽  
1H And 13C Nmr ◽  
One Dimensional ◽  
Linkage Analyses ◽  
Yariv Reagent ◽  
Native Tree ◽  
High Level

A gum that exudes from the wounded trunk of the New Zealand native tree Meryta sinclairii has been isolated. The gum was completely precipitated by the β-glucosyl Yariv reagent and was thus determined to be an arabinogalactan-protein (AGP). It contained >95% w/w carbohydrate and only 2% w/w protein with a high level of hydroxyproline. SEC-MALLS showed that the gum had a weight-average molecular weight of 4.45×106Da compared with 6.02×105Da for gum arabic. Constituent sugar and linkage analyses were consistent with polymers comprised of a highly branched backbone of 1,3-linked galactopyranosyl (Galp) residues, with side-chains made up of arabinofuranose- (Araf) containing oligosaccharides, terminated variously by rhamnopyranosyl (Rhap), arabinopyranosyl (Arap), Galp and glucuronopyranosyl (GlcpA) residues. Analysis by one-dimensional and two-dimensional 1H and 13C NMR experiments confirmed the linkage analyses. The structure of the gum is discussed in comparison with the structure of gum arabic and other AGPs. © 2003 Elsevier Science Ltd. All rights reserved.

Molecular, rheological and physicochemical characterisation of puka gum, an arabinogalactan-protein extracted from the Meryta sinclairii tree

2020 ◽  
Author(s):  
MSM Wee ◽  
Ian Sims ◽  
KKT Goh ◽  
L Matia-Merino
Keyword(s):  
Hydrodynamic Radius ◽  
Average Molecular Weight ◽  
Gum Arabic ◽  
Water Soluble ◽  
Arabinogalactan Protein ◽  
Type Ii ◽  
Weight Average Molecular Weight ◽  
Soluble Polysaccharide ◽  
Physicochemical Characterisation ◽  
Increasing Temperature

© 2019 Elsevier Ltd A water-soluble polysaccharide (type II arabinogalactan-protein) extracted from the gum exudate of the native New Zealand puka tree (Meryta sinclairii), was characterised for its molecular, rheological and physicochemical properties. In 0.1 M NaCl, the weight average molecular weight (Mw) of puka gum is 5.9 × 106 Da with an RMS radius of 56 nm and z-average hydrodynamic radius of 79 nm. The intrinsic viscosity of the polysaccharide is 57 ml/g with a coil overlap concentration 15% w/w. Together, the shape factor, p, of 0.70 (exponent of RMS radius vs. hydrodynamic radius), Smidsrød-Haug's stiffness parameter B of 0.031 and Mark-Houwink exponent α of 0.375 indicate that the polysaccharide adopts a spherical conformation in solution, similar to gum arabic. The pKa is 1.8. The polysaccharide exhibits a Newtonian to shear-thinning behaviour from 0.2 to 25% w/w. Viscosity of the polysaccharide (1 s−1) decreases with decreasing concentration, increasing temperature, ionic strength, and at acidic pH.

Extracellular polysaccharides from suspension cultures of Nicotiana plumbaginifolia

2020 ◽  
Author(s):  
Ian Sims ◽  
A Bacic
Keyword(s):  
Uronic Acid ◽  
Cell Suspension Cultures ◽  
Anion Exchange Chromatography ◽  
Nicotiana Plumbaginifolia ◽  
Suspension Cultures ◽  
Exchange Chromatography ◽  
Arabinogalactan Protein ◽  
Extracellular Polysaccharides ◽  
Selective Precipitation ◽  
The Individual

The soluble polymers secreted by cell-suspension cultures of Nicotiana plumbaginifolia contained 78% carbohydrate, 6% protein and 4% inorganic material. The extracellular polysaccharides were separated into three fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7 and the individual polysaccharides in each fraction were then isolated by selective precipitation and enzymic treatment. Monosaccharide and linkage compositions were determined for each polysaccharide after reduction of uronic acid residues and the degree of esterification of the various uronic acid residues in each polysaccharide was determined concurrently with the linkage types. Six components were identified: an arabinoxyloglucan (comprising 34% of the total polysaccharide) and a galactoglucomannan (15%) in the unbound neutral fraction, a type II arabinogalactan (an arabinogalactan-protein, 11%) and an acidic xylan (3%) in the first bound fraction, and an arabinoglucuronomannan (11%) and a galacturonan (26%) in the second bound fraction. © 1995.

scholarly journals High level expression of transgenes by use of 5^|^#8242;-untranslated region of the Arabidopsis thaliana arabinogalactan-protein 21 gene in dicotyledons

2012 ◽  
Vol 29 (3) ◽  
pp. 319-322 ◽  
Author(s):  
Takeshi Matsui ◽  
Hideyuki Matsuura ◽  
Kazutoshi Sawada ◽  
Eiji Takita ◽  
Satoko Kinjo ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Untranslated Region ◽  
Arabinogalactan Protein ◽  
High Level Expression ◽  
High Level

scholarly journals Characterization of the hydroxyproline-rich protein core of an arabinogalactan-protein secreted from suspension-cultured Lolium multiflorum (Italian ryegrass) endosperm cells

1989 ◽  
Vol 264 (3) ◽  
pp. 857-862 ◽  
Author(s):  
P A Gleeson ◽  
M McNamara ◽  
R E H Wettenhall ◽  
B A Stone ◽  
G B Fincher
Keyword(s):  
Polyclonal Antibodies ◽  
Lolium Multiflorum ◽  
Italian Ryegrass ◽  
Arabinogalactan Protein ◽  
Myeloma Protein ◽  
Protein Core ◽  
Liquid Suspension ◽  
Peptide Component ◽  
Immunodominant Epitopes

An arabinogalactan-protein (AGP) purified from the filtrate of liquid-suspension-cultured Italian-ryegrass (Lolium multiflorum) endosperm cells by affinity chromatography on myeloma protein J539-Sepharose was deglycosylated with trifluoromethanesulphonic acid to remove polysaccharide chains that are covalently associated with hydroxyproline residues in the peptide component of the proteoglycan. The protein core, which accounts for less than 10% (w/w) of the intact proteoglycan, was purified by h.p.l.c. It has an apparent Mr of 35,000, but reacts very poorly with both Coomassie Brilliant Blue R and silver stains. Amino-acid-sequence analysis of the N-terminus of the h.p.l.c.-purified protein core and of tryptic peptides generated from the unpurified protein reveals a high content of hydroxyproline and alanine. These are sometimes arranged in short (Ala-Hyp) repeat sequences of up to six residues. Polyclonal antibodies raised against the protein core do not cross-react with native AGP, the synthetic peptide (Ala-Hyp)4, poly-L-hydroxyproline or poly-L-proline. The results suggest that the polysaccharide chains in the native AGP render the protein core of the proteoglycan inaccessible to the antibodies and that the immunodominant epitopes include domains of the protein other than those rich in Ala-Hyp repeating units.

Fine structure of the arabinogalactan-protein from Lolium multiflorum

1987 ◽  
Vol 162 (1) ◽  
pp. 85-93 ◽  
Author(s):  
Antony Bacic ◽  
Shirley C. Churms ◽  
Alistair M. Stephen ◽  
Peter B. Cohen ◽  
Geoffrey B. Fincher
Keyword(s):  
Fine Structure ◽  
Lolium Multiflorum ◽  
Arabinogalactan Protein
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