Isoelectric focusing of peroxidases released from soybean hypocotyl cell walls by Sclerotium rolfsii culture filtrate

1974 ◽  
Vol 52 (9) ◽  
pp. 2037-2040 ◽  
Author(s):  
C. R. Curtis ◽  
Neal M. Barnett
Keyword(s):  
Cell Wall ◽  
Isoelectric Focusing ◽  
Culture Filtrate ◽  
Esterase Activity ◽  
Cell Walls ◽  
Sclerotium Rolfsii ◽  
Disc Electrophoresis ◽  
Fungal Culture ◽  
Cell Wall Proteins ◽  
Isoelectric Points

Bound soybean hypocotyl cell wall proteins can be released from the cell wall by incubation with a culture filtrate of the fungal pathogen Sclerotium rolfsii. The released cell wall proteins were then separated by disc electrophoresis or isoelectric-focusing techniques in polyacrylamide gel columns and tested for peroxidase and esterase activity. Released cell wall proteins showed considerable peroxidase multiplicity, which was totally absent in the separation of fungal culture filtrate. Peroxidase patterns of some released cell wall peroxidases were similar to soybean cytoplasmic peroxidase patterns. In separate experiments, esterase bands were detected in the released cell wall proteins, but this was probably due to esterase activity of the fungal culture filtrate. Cytoplasmic esterases exhibited much higher isoelectric points than cell wall and culture filtrate esterases, which were concentrated between pH 3 and 6.

scholarly journals Co-operative action by endo- and exo-β-(1→3)-glucanases from parasitic fungi in the degradation of cell-wall glucans of Sclerotinia sclerotiorum (Lib.) de Bary

1974 ◽  
Vol 140 (1) ◽  
pp. 47-55 ◽  
Author(s):  
David Jones ◽  
Alex. H. Gordon ◽  
John S. D. Bacon
Keyword(s):  
Cell Wall ◽  
Sclerotinia Sclerotiorum ◽  
Culture Filtrate ◽  
Cell Walls ◽  
Trichoderma Viride ◽  
Major Constituent ◽  
Coniothyrium Minitans ◽  
Parasitic Fungi ◽  
Complete Breakdown

1. Two fungi, Coniothyrium minitans Campbell and Trichoderma viride Pers. ex Fr., were grown on autoclaved crushed sclerotia of the species Sclerotinia sclerotiorum, which they parasitize. 2. in vitro the crude culture filtrates would lyse walls isolated from hyphal cells or the inner pseudoparenchymatous cells of the sclerotia, in which a branched β-(1→3)-β-(1→6)-glucan, sclerotan, is a major constituent. 3. Chromatographic fractionation of the enzymes in each culture filtrate revealed the presence of several laminarinases, the most active being an exo-β-(1→3)-glucanase, known from previous studies to attack sclerotan. Acting alone this brought about a limited degradation of the glucan, but the addition of fractions containing an endo-β-(1→3)-glucanase led to almost complete breakdown. A similar synergism between the two enzymes was found in their lytic action on cell walls. 4. When acting alone the endo-β-(1→3)-glucanase had a restricted action, the products including a trisaccharide, tentatively identified as 62-β-glucosyl-laminaribiose. 5. These results are discussed in relation to the structure of the cell walls and of their glucan constituents.

scholarly journals Di-isodityrosine, a novel tetrametric derivative of tyrosine in plant cell wall proteins: a new potential cross-link

1996 ◽  
Vol 315 (1) ◽  
pp. 323-327 ◽  
Author(s):  
Jeffrey D. BRADY ◽  
Ian H. SADLER ◽  
Stephen C. FRY
Keyword(s):  
Cell Culture ◽  
Cell Wall ◽  
Amino Acid ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cell Wall Proteins ◽  
Cross Link ◽  
Nmr Spectrometry

A novel amino acid, di-isodityrosine, has been isolated from hydrolysates of cell walls of tomato cell culture. Analysis by UV spectrometry, partial derivatization with 2,4-dinitrofluorobenzene and mass and NMR spectrometry show that the compound is composed to two molecules of isodityrosine, joined by a biphenyl linkage. The possible reactions involved in the formation of this molecule in vivo are discussed, as is the possibility that it could form an interpolypeptide linkage between cell wall proteins such as extensin, and hence aid in the insolubilization of the protein in the wall.

scholarly journals Abscission Layer Formation as a Resistance Response of Peruvian Apple Cactus Against Glomerella cingulata

2002 ◽  
Vol 92 (9) ◽  
pp. 964-969 ◽  
Author(s):  
Young Ho Kim ◽  
Kwang-Hyung Kim
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Cell Walls ◽  
Light And Electron Microscopy ◽  
Middle Lamella ◽  
Fungal Culture ◽  
Glomerella Cingulata ◽  
Abscission Layer ◽  
Resistance Response ◽  
Initial Cell

Stem disks from 2-year-old cacti Cereus tetragonus (susceptible) and C. peruvianus (resistant) were inoculated in the center (pith) with Glomerella cingulata isolated from Colletotrichum stem rot in three-angled cacti. The susceptible cactus became extensively colonized, whereas colonization was limited to a small area in the resistant cactus. The resistant cactus formed prominent abscission layers (ALs) in parenchyma internal to the inoculation site. Ethanol extracts of the fungal culture also stimulated AL formation in the resistant cactus. Initial cell division followed at 2 to 4 days after treatment, and layering of multiple cells at 7 days after treatment. After 10 days, the outer layers were sometimes sloughed from the inner layers. No AL formation was induced in susceptible C. tetragonus treated with ethanol extract or in untreated control cacti. Light and electron microscopy revealed that initial cell division occurred by cell wall formation, and that an additional cell wall was layered in pre-existing parenchyma cells without ordinary cell division. Later, separation layers formed in ALs where inner cell walls appeared to be thickened secondarily, and the cell walls and middle lamella within the layer dissolved. These results suggest that AL formation in the resistant cactus is induced by fungal metabolites, and that it serves as a histological barrier against anthracnose pathogens.

Composition and properties of the cell wall of Methanospirillum hungatii

1980 ◽  
Vol 26 (2) ◽  
pp. 115-120 ◽  
Author(s):  
G. D. Sprott ◽  
R. C. McKellar
Keyword(s):  
Amino Acids ◽  
Cell Wall ◽  
Cell Walls ◽  
Weight Fraction ◽  
Dry Weight ◽  
Wall Material ◽  
Bond Breaking ◽  
High Molecular Weight Fraction ◽  
Cell Wall Proteins ◽  
Isolated Cell

Dithiothreitol reacted, at pH 9.0, with the isolated cell walls of Methanospirillum hungatii, to release about 23% of the cell wall dry weight as a high molecular weight fraction (> 0.5 million daltons). Untreated walls consisted of 70% amino acids, 11% lipid, and 6.6% carbohydrate. Sugars were identified as rhamnose, ribose, glucose, galactose, and mannose. The wall material that was released contained only 47% amino acids and was enriched in lipid, glucose, and phosphate. These results support data from electron micrographs, showing the localized release of cell wall material by the disulfide bond-breaking reagent at alkaline pH. In amino acid composition the untreated walls did not differ greatly from the material released by dithiothreitol, but differed considerably from the walls of another strain of M. hungatii. The ratios of the amino acids found in the cell wall proteins of several archaebacteria and of Bacillus cereus spore coats were similar.

Release of peroxidase from soybean hypocotyl cell walls by Sclerotium rolfsii culture filtrates

1974 ◽  
Vol 52 (1) ◽  
pp. 265-271 ◽  
Author(s):  
Neal M. Barnett
Keyword(s):  
Culture Filtrate ◽  
Peroxidase Activity ◽  
Cell Walls ◽  
Hydrolytic Enzymes ◽  
Sclerotium Rolfsii ◽  
Release Reaction ◽  
Culture Filtrates ◽  
Ph Range

Up to 24% of the peroxidase of purified cell walls of soybean hypocotyls was released by incubation of cell walls with hydrolytic enzymes secreted by the fungus Sclerotium rolfsii. This estimate is based on comparison of peroxidase activity recovered in the medium with peroxidase activity in unincubated cell walls, estimated by a new assay. The peroxidase-release reaction occurs at 0 °C at half the rate at 30 °C. The peroxidase-release reaction occurs almost equally fast in the pH range of 3.5 to 8.0. The release of peroxidase from cell walls cannot be attributed solely to arabanase, polygalacturonase, or cellulase in the culture filtrate, although on Sephadex G-75 chromatography these activities overlap the peroxidase-releasing activity. Culture filtrate released less than 5% of the hydroxyproline protein of the cell walls.

scholarly journals Identification and characterization of plant glycerophosphodiester phosphodiesterase

2004 ◽  
Vol 379 (3) ◽  
pp. 601-607 ◽  
Author(s):  
Benoît van der REST ◽  
Norbert ROLLAND ◽  
Anne-Marie BOISSON ◽  
Myriam FERRO ◽  
Richard BLIGNY ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Cell Walls ◽  
Expression System ◽  
Gel Filtration ◽  
Cell Suspension Cultures ◽  
Cell Wall Proteins ◽  
Ammonium Sulphate Precipitation ◽  
Carrot Cell ◽  
Glycerophosphodiester Phosphodiesterase

GPX-PDE (glycerophosphodiester phosphodiesterase; EC 3.1.4.46) is a relatively poorly characterized enzyme that catalyses the hydrolysis of various glycerophosphodiesters (glycerophosphocholine, glycerophosphoethanolamine, glycerophosphoglycerol, glycerophosphoserine and bis-glycerophosphoglycerol), releasing sn-glycerol 3-phosphate and the corresponding alcohol. In a previous study, we demonstrated the existence of a novel GPX-PDE in the cell walls and vacuoles of plant cells. Since no GPX-PDE had been identified in any plant organism, the purification of GPX-PDE from carrot cell walls was attempted. After extraction of cell wall proteins from carrot cell suspension cultures with CaCl2, GPX-PDE was purified up to 2700-fold using, successively, ammonium sulphate precipitation, gel filtration and concanavalin A–Sepharose. Internal sequence analysis of a 55 kDa protein identified in the extract following 2700-fold purification revealed strong similarity to the primary sequence of GLPQ, a bacterial GPX-PDE. To confirm the identity of plant GPX-PDE, an Arabidopsis thaliana cDNA similar to that encoding the bacterial GPX-PDE was cloned and overexpressed in a bacterial expression system, and was used to raise antibodies against the putative Arabidopsis thaliana GPX-PDE. Immunochemical assays performed on carrot cell wall proteins extracted by CaCl2 treatment showed a strong correlation between GPX-PDE activity and detection of the 55 kDa protein, validating the identity of the plant GPX-PDE. Finally, various properties of the purified enzyme were investigated. GPX-PDE is a multimeric enzyme, specific for glycerophosphodiesters, exhibiting a Km of 36 µM for glycerophosphocholine and active within a wide pH range (from 4 to 10). Since these properties are similar to those of GLPQ, the bacterial GPX-PDE, the similarities between plant and bacterial enzymes are also discussed.

scholarly journals Plant Cell Wall Proteins and Development

2020 ◽  
Vol 21 (8) ◽  
pp. 2731
Author(s):  
Elisabeth Jamet ◽  
Christophe Dunand
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cell Communication ◽  
Cell Wall Proteins ◽  
Plant Cell Walls

Plant cell walls surround cells and provide both external protection and a means of cell-to-cell communication [...]

scholarly journals Sed1p Is a Major Cell Wall Protein ofSaccharomyces cerevisiae in the Stationary Phase and Is Involved in Lytic Enzyme Resistance

1998 ◽  
Vol 180 (13) ◽  
pp. 3381-3387 ◽  
Author(s):  
Hitoshi Shimoi ◽  
Hiroshi Kitagaki ◽  
Hisanobu Ohmori ◽  
Yuzuru Iimura ◽  
Kiyoshi Ito
Keyword(s):  
Cell Wall ◽  
Stationary Phase ◽  
Cell Walls ◽  
Secretory Pathway ◽  
Signal Sequence ◽  
Sodium Dodecyl ◽  
Cell Wall Protein ◽  
Lytic Enzyme ◽  
Cell Wall Proteins ◽  
Structural Cell

ABSTRACT A 260-kDa structural cell wall protein was purified from sodium dodecyl sulfate-treated cell walls of Saccharomyces cerevisiae by incubation with Rarobacter faecitabidusprotease I, which is a yeast-lytic enzyme. Amino acid sequence analysis revealed that this protein is the product of the SED1 gene.SED1 was formerly identified as a multicopy suppressor oferd2, which encodes a protein involved in retrieval of luminal endoplasmic reticulum proteins from the secretory pathway. Sed1p is very rich in threonine and serine and, like other structural cell wall proteins, contains a putative signal sequence for the addition of a glycosylphosphatidylinositol anchor. However, the fact that Sed1p, unlike other cell wall proteins, has six cysteines and seven putative N-glycosylation sites suggests that Sed1p belongs to a new family of cell wall proteins. Epitope-tagged Sed1p was detected in a β-1,3-glucanase extract of cell walls by immunoblot analysis, suggesting that Sed1p is a glucanase-extractable cell wall protein. The expression of Sed1p mRNA increased in the stationary phase and was accompanied by an increase in the Sed1p content of cell walls. Disruption of SED1 had no effect on exponentially growing cells but made stationary-phase cells sensitive to Zymolyase. These results indicate that Sed1p is a major structural cell wall protein in stationary-phase cells and is required for lytic enzyme resistance.

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