Search results for "cell wall"
showing 10 items of 226 documents
Characterization of cell wall proteins from yeast and mycelial cells of Candida albicans by labelling with biotin: Comparison with other techniques
1992
Candida albicans ATCC 26555 blastoconidia and blastoconidia bearing germ tubes were metabolically labelled by incubating the cells with 14C-labelled protein hydrolysate and were subsequently tagged with biotin. Double-labelled (radioactive and biotinylated) cell wall proteins and glycoproteins were extracted from intact cells of both growth forms by treatment with 2-mercaptoethanol (beta ME) and with beta-glucanases (Zymolyase) after treatment with beta ME. The beta ME- and Zymolyase-extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotted (immunoblotted) to nitrocellulose paper. Polyacrylamide gels were stained with Coomassie blue and process…
Preliminary characterization of the material released to the culture medium by Candida albicans yeast and mycelial cells.
1995
Culture filtrate concentrates were obtained from Candida albicans yeast and mycelial cells grown in the presence of 14C-protein hydrolysate for radioactive labeling of cellular polypeptides. Both growth forms released to the medium minor but significant amounts of proteinaceous materials. The analysis of culture filtrate concentrates by means of SDS-polyacrylamide gel electrophoresis and fluorography revealed a similar and complex electrophoretic pattern, though some qualitative and quantitative differences between samples obtained from yeast and mycelial cells were observed. Materials released, mostly composed of mannoproteins as shown by their affinity towards concanavalin A, presented (i…
Experimental virus evolution reveals a role of plant microtubule dynamics and TORTIFOLIA1/SPIRAL2 in RNA trafficking.
2014
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Overexpression of a cell wall enzyme reduces xyloglucan depolymerization and softening of transgenic tomato fruits.
2010
Xyloglucan xyloglucosyltransferase/endohydrolase (XTHs: EC 2.4.1.207 and/or EC 3.2.1.151) has been proposed to have a dual role integrating newly secreted xyloglucan chains into an existing wall-bound xyloglucan and restructuring existing cell wall material by catalyzing transglucosylation between previously wall bound xyloglucan molecules. In this work we generated transgenic tomatoes with altered levels of an XTH gene. These transgenic fruits showed significant overexpression of the XTH proteins in comparison with the wild type. Specific XET activity was approximately 4.33 fold higher in the transgenic fruits compared with the wild type fruits, although in both cases the activity decrease…
Cloning and characterization of the phenylalanyl-tRNA synthetase β subunit gene fromCandida albicans
1998
A Candida albicans expression library was constructed from RNA isolated from regenerating protoplasts. A 1.4-kb cDNA clone was used to isolate a genomic fragment. Sequence analysis revealed an open reading frame of 593 amino acids with an overall identity of 63.6% with the phenylalanyl-tRNA synthetase beta subunit (FRS1) of Saccharomyces cerevisiae. We named it CaFRS1. It is located in a single copy in chromosome R, SfiI fragment M. Its expression showed a decrease during the cell wall regeneration process in protoplasts of both yeast and mycelial cells of C. albicans, suggesting its requirement thereof in initial steps of the cell wall synthesis.
Sedimentation properties of chitosomal chitin synthetase from the wild-type strain and the 'slime' variant of Neurospora crassa.
1989
Marked differences in the pattern of sedimentation of cellular structures were observed after isopycnic centrifugation of crude cell-free preparations from the Neurospora crassa wall-less 'slime' variant and mycelial wild-type strain. Kinetic studies of particle sedimentation showed that the various types of subcellular components, as revealed by turbidity, UV absorption, polypeptide patterns, and chitin synthetase activity determinations, sediment independently of one another. An important feature was the finding that chitin synthetase from 'slime' peaked at a median specific gravity of 1.1201 +/- 0.0036, whereas that from wild-type strain sedimented at a higher buoyant density (specific g…
Reaggregation and binding of cell wall proteins from Candida albicans to structural polysaccharides
1998
Urea or hot sodium dodecyl sulphate extracted a significant amount of the same proteins from the matrix of the cell wall of the yeast form and mycelial cells of Candida albicans. Gel filtration analysis of the urea-extracted proteins revealed that they occurred in the form of large complexes which were unaffected by up to 8 M urea. Among them, proteins en route to becoming covalently associated within the wall scaffold were identified by their reaction with specific antibodies. When urea was removed by dialysis, some of these proteins specifically reassociated into large aggregates which bound strongly with ConA, whereas others remained soluble in smaller associated products. The ability of…
Characterization of a disulphide-bound Pir-cell wall protein (Pir-CWP) ofYarrowia lipolytica
2003
In this work we have studied the disulphide-bound group of cell wall mannoproteins of Yarrowia lipolytica and Candida albicans. In the case of Y. lipolytica, SDS-PAGE analysis of the beta-mercaptoethanol-extracted material from the purified cell walls of the yeast form, showed the presence of a main polypeptide of 45 kDa and some minor bands in the 100-200 kDa range. This pattern of bands is similar to that obtained in identical extracts in Saccharomyces cerevisiae (Moukadiri et al., 1999), and besides, all these bands cross-react with an antibody raised against beta-mercaptoethanol-extracted material from the purified cell walls of S. cerevisiae, suggesting that the 45 kDa band could be th…
Drying enhances immunoactivity of spent brewer's yeast cell wall β-D-glucans.
2015
Due to immunological activity, microbial cell wall polysaccharides are defined as 'biological response modifiers' (BRM). Cell walls of spent brewer's yeast also have some BRM activity. However, up to date there is no consensus on the use of spent brewer's yeast D-glucan as specific BRM in humans or animals. The aim of this paper is to demonstrate the potential of spent brewer's yeast β-D-glucans as BRM, and drying as an efficient pretreatment to increase β-D-glucan's immunogenic activity. Our results revealed that drying does not change spent brewer's yeast biomass carbohydrate content as well as the chemical structure of purified β-D-glucan. However, drying increased purified β-D-glucan TN…
Transglutaminase activity is involved in Saccharomyces cerevisiae wall construction
2002
Transglutaminase activity, which forms the interpeptidic cross-link N(epsilon)-(gamma-glutamyl)-lysine, was demonstrated in cell-free extracts of Saccharomyces cerevisiae by incorporation of [(14)C]lysine into an exogenous acceptor, N,N'-dimethylcasein. Higher levels of the activity were present in the cell wall, which also contained endogenous acceptors. The enzyme activity in the wall was inhibited by cystamine, a known inhibitor of transglutaminase, and by EDTA, indicating a cation-dependent activity. After the endogenous wall acceptors were labelled radioactively by transglutaminase, extraction with SDS solubilized about 50% of the total radioactivity, while Zymolyase and chitinase each…