scholarly journals Cell Wall Growth and Modulation Dynamics in a Model Unicellular Green Alga—Penium margaritaceum: Live Cell Labeling with Monoclonal Antibodies

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
David S. Domozych ◽  
Hannah Brechka ◽  
Alicia Britton ◽  
Marc Toso
Keyword(s):  
Cell Wall ◽  
Monoclonal Antibodies ◽  
Green Alga ◽  
Fluorescent Labeling ◽  
Land Plants ◽  
Cell Labeling ◽  
Live Cells ◽  
Specific Cell ◽  
Cell Wall Degrading Enzymes ◽  
Cell Wall Polymers

Penium margaritaceum is a unicellular charophycean green alga that possesses cell wall polymers similar to those of land plants. Several wall macromolecules of this alga are recognized by monoclonal antibodies specific for wall polymer epitopes of land plants. Immunofluorescence protocols using these antibodies may be employed to label specific cell wall constituents of live cells. Fluorescent labeling persists for several days, and this attribute allows for tracing of wall epitopes in both long- and short-term studies of cell development. Quantitative analysis of surface area covered by cell wall polymers is also easily performed. We show that significant cell expansion caused by incubation of cells in low levels of osmotically active agents like mannitol, glucose, or sucrose results from the inability of cells to undergo cytokinesis but does not result in significant changes to the amount of new cell wall. We also demonstrate that cells can be maintained for long periods of time in culture medium supplemented with specific cell wall-degrading enzymes where notable changes to wall infrastructure occur. These results demonstrate the great potential value of Penium in elucidating fundamental events during cell wall synthesis and modulation in plant cells.

The Cell Wall Polymers of the Charophycean Green Alga Chara corallina: Immunobinding and Biochemical Screening

2010 ◽  
Vol 171 (4) ◽  
pp. 345-361 ◽  
Author(s):  
David S. Domozych ◽  
Iben Sørensen ◽  
Filomena A. Pettolino ◽  
Anthony Bacic ◽  
William G. T. Willats
Keyword(s):  
Cell Wall ◽  
Green Alga ◽  
Chara Corallina ◽  
Biochemical Screening ◽  
Cell Wall Polymers

scholarly journals Identification of a Lipoteichoic Acid Glycosyltransferase Enzyme Reveals that GW-Domain-Containing Proteins Can Be Retained in the Cell Wall of Listeria monocytogenes in the Absence of Lipoteichoic Acid or Its Modifications

2016 ◽  
Vol 198 (15) ◽  
pp. 2029-2042 ◽  
Author(s):  
Matthew G. Percy ◽  
Eleni Karinou ◽  
Alexander J. Webb ◽  
Angelika Gründling
Keyword(s):  
Cell Wall ◽  
Listeria Monocytogenes ◽  
Bacterial Genome ◽  
Lipoteichoic Acid ◽  
Specific Cell ◽  
Cell Wall Proteins ◽  
Wall Teichoic Acid ◽  
Content Type ◽  
Cell Wall Polymer ◽  
Cell Wall Polymers

ABSTRACTListeria monocytogenesis a foodborne Gram-positive bacterial pathogen, and many of its virulence factors are either secreted proteins or proteins covalently or noncovalently attached to the cell wall. Previous work has indicated that noncovalently attached proteins with GW (glycine-tryptophan) domains are retained in the cell wall by binding to the cell wall polymer lipoteichoic acid (LTA). LTA is a glycerol phosphate polymer, which is modified inL. monocytogeneswith galactose andd-alanine residues. We identified Lmo0933 as the cytoplasmic glycosyltransferase required for the LTA glycosylation process and renamed the protein GtlA, forglycosyltransferaseLTAA. UsingL. monocytogenesmutants lacking galactose ord-alanine modifications or the complete LTA polymer, we show that GW domain proteins are retained within the cell wall, indicating that other cell wall polymers are involved in the retention of GW domain proteins. Further experiments revealed peptidoglycan as the binding receptor as a purified GW domain fusion protein can bind toL. monocytogenescells lacking wall teichoic acid (WTA) as well as purified peptidoglycan derived from a wild-type or WTA-negative strain. With this, we not only identify the first enzyme involved in the LTA glycosylation process, but we also provide new insight into the binding mechanism of noncovalently attached cell wall proteins.IMPORTANCEOver the past 20 years, a large number of bacterial genome sequences have become available. Computational approaches are used for the genome annotation and identification of genes and encoded proteins. However, the function of many proteins is still unknown and often cannot be predicted bioinformatically. Here, we show that the previously uncharacterizedListeria monocytogenesgenelmo0933likely codes for a glycosyltransferase required for the decoration of the cell wall polymer lipoteichoic acid (LTA) with galactose residues. UsingL. monocytogenesmutants lacking LTA modifications or the complete polymer, we show that specific cell wall proteins, often associated with virulence, are retained within the cell wall, indicating that additional cell wall polymers are involved in their retention.

Bacterial surface antigen-specific monoclonal antibodies used to detect beer spoilage pediococci

1999 ◽  
Vol 45 (8) ◽  
pp. 670-677 ◽  
Author(s):  
Michael S Whiting ◽  
W M Ingledew ◽  
Sun Y Lee ◽  
Barry Ziola
Keyword(s):  
Cell Wall ◽  
Monoclonal Antibodies ◽  
Surface Antigen ◽  
Surface Antigens ◽  
Bacterial Surface ◽  
Cell Wall Polymers ◽  
Good Potential ◽  
Protease Treatment ◽  
Beer Spoilage

Fourteen monoclonal antibodies (Mabs) were isolated that react with surface antigens of Pediococcus beer spoilage organisms, including P. damnosus, P. pentosaceous, P. acidilactici, and unspeciated isolates. Immunoblotting, enzyme immunoassays (EIAs) of protease- and neuraminidase-treated surface antigen extracts, carbohydrate competition EIAs, and cardiolipin EIAs were used to characterize the bacterial antigens involved in Mab binding. Antigen stability in situ was tested by protease treatment or surface antigen extraction of washed bacteria. In most cases, the Mabs bind to Pediococcus surface antigens that appear to be covalently bound cell wall polymers resistant to alteration or removal from the bacterial surface. These bacterial surface antigen reactive Mabs show good potential for rapid, sensitive, and specific immunoassay detection of Pediococcus beer spoilage organisms.Key words: beer spoilage organism, immunoassay, monoclonal antibodies, Pediococcus, surface antigens.

scholarly journals Cell Wall Degradation in Irradiated Tomato Fruit

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 815B-815
Author(s):  
N. El-Assi ◽  
D.J. Huber ◽  
J.K. Brecht
Keyword(s):  
Cell Wall ◽  
Tomato Fruit ◽  
Selective Extraction ◽  
Specific Cell ◽  
Neutral Sugar ◽  
Cell Wall Degradation ◽  
X Ray ◽  
Green Fruit ◽  
Cell Wall Polymers

The irradiation of harvested fruit is typically accompanied by excessive tissue softening, a process that is not well understood. In this study, we examined the role of specific cell wall polymers and the extent of general cell wall degradation and softening in irradiated tomato fruit. `Sunny' tomato fruit at mature-green and pink stages were subjected to X-ray radiation at 0, 83, and 156 Krad. Immediate softening was noted for both maturation classes, although some postirradiation recovery was evident in green fruit. Pectic polymers of both mature-green and pink fruit exhibited depolymerization and altered neutral sugar profiles in response to irradiation. Pectins, either as components of total ethanol-insoluble solids (EIS), purified by selective extraction, or of commercial origin were similarly affected by irradiation. Cellulose preparations were unaffected by irradiation. The data demonstrate that the effect of irradiation on the cell wall exhibits specificity, can occur nonenzymatically, and does not require initiating adducts of cytosolic origin.

scholarly journals The distribution of cell wall polymers during antheridium development and spermatogenesis in the Charophycean green alga, Chara corallina

2009 ◽  
Vol 104 (6) ◽  
pp. 1045-1056 ◽  
Author(s):  
David S. Domozych ◽  
Iben Sørensen ◽  
William G. T. Willats
Keyword(s):  
Cell Wall ◽  
Green Alga ◽  
Chara Corallina ◽  
Cell Wall Polymers

scholarly journals Valorization of sugarcane bagasse by chemical pretreatment and enzyme mediated deconstruction

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Vihang S. Thite ◽  
Anuradha S. Nerurkar
Keyword(s):  
Cell Wall ◽  
Sugarcane Bagasse ◽  
Structural Changes ◽  
Enzymatic Saccharification ◽  
Dry Weight ◽  
Gravimetric Analysis ◽  
Chemical Pretreatment ◽  
Cell Wall Degrading Enzymes ◽  
First Time ◽  
Soluble Components

Abstract After chemical pretreatment, improved amenability of agrowaste biomass for enzymatic saccharification needs an understanding of the effect exerted by pretreatments on biomass for enzymatic deconstruction. In present studies, NaOH, NH4OH and H2SO4 pretreatments effectively changed visible morphology imparting distinct fibrous appearance to sugarcane bagasse (SCB). Filtrate analysis after NaOH, NH4OH and H2SO4 pretreatments yielded release of soluble reducing sugars (SRS) in range of ~0.17–0.44%, ~0.38–0.75% and ~2.9–8.4% respectively. Gravimetric analysis of pretreated SCB (PSCB) biomass also revealed dry weight loss in range of ~25.8–44.8%, ~11.1–16.0% and ~28.3–38.0% by the three pretreatments in the same order. Release of soluble components other than SRS, majorly reported to be soluble lignins, were observed highest for NaOH followed by H2SO4 and NH4OH pretreatments. Decrease or absence of peaks attributed to lignin and loosened fibrous appearance of biomass during FTIR and SEM studies respectively further corroborated with our observations of lignin removal. Application of commercial cellulase increased raw SCB saccharification from 1.93% to 38.84%, 25.56% and 9.61% after NaOH, H2SO4 and NH4OH pretreatments. Structural changes brought by cell wall degrading enzymes were first time shown visually confirming the cell wall disintegration under brightfield, darkfield and fluorescence microscopy. The microscopic evidence and saccharification results proved that the chemical treatment valorized the SCB by making it amenable for enzymatic saccharification.

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