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PubMed1968

The presence of ribonucleic acid in the cell walls of higher plants.

A method for isolating extensively purified cell walls from higher plants is described; the preparations contain no detectable chloroplast or nuclear material and the protein content (2-5% of the dry wt. of walls) indicates that there is little contamination with cytoplasm. Incubation of purified cell walls with 0.3n-potassium hydroxide for 17hr. at 37 degrees liberates ribonucleotides, which can be purified by adsorption on charcoal and by ion-exchange chromatography. Ribonucleotides are also liberated by incubating the walls with ribonuclease, but not with deoxyribonuclease. The RNA content varies from 0.5 to 6mg./g. dry wt. of walls, depending on the nature and age of the tissue, and at 3mg./g. dry wt. of walls accounts for about 7% of the total RNA of the tissue. Less than 0.2% of the RNA of the walls is due to the presence of bacteria in the preparation. The base composition of the cell-wall RNA is identical with that of ribosomal RNA.

Age Factors↗
PubMed1980

Regulation of cell wall mycolic acid biosynthesis in acid-fast bacteria. I. Temperature-induced changes in mycolic acid molecular species and related compounds in Mycobacterium phlei.

Molecular species of two major subclasses of mycolic acids from Mycobacterium phlei, alpha-mycolic acids (M1) and dicarboxy mycolic acids (M3), were separated gas-chromatographically and identified mass-spectrometrically. The mycolic acid compositions of extractable and cell wall bound lipids were markedly influenced by growth temperature. Increasing growth temperature from 20 degrees C to 50 degrees C resulted in an increase in longer chain species of both mycolisc acid subclasses with a concomitant decrease in shorter chain homologues. The most abundant molecular species were C76 and C58 of M1 and M3 in the 20 degrees C grown cells, while the 50 degrees C grown cells contained C80 in M1 and C62 in M3, most abundantly. Changes in mycolic acid composition occurred rapidly after growth temperature was raised from 20 degrees C to 50 degrees C with an increase in C62 and a concomitant decrease in C58. Mass fragmentographic analysis revealed that an increase in total carbon numbers of mycolic acids was caused by the elongation of straight chain alkyl unit, without any changes in alpha-branch. Changes in the molecular species composition of secondary alcohols presumably derived from the ester mycolic acids were also observed and an increase in longer species (C20-ol-2) with a concomitant decrease in shorter ones (C18-ol-2) was noted as the temperature rose. An increase in the growth temperature also resulted in a decrease in unsaturated fatty acids in extractable lipids. These observations suggest that mycobacteria alter the molecular species composition of mycolic acid subclasses and phospholipids, in response to growth temperature, to maintain a suitable membrane function.

Cell Wall↗
PubMed1975

Characterization of two cell-envelope fractions from chemotrophically grown Rhodospirillum rubrum.

Two cell-envelope fractions were isolated from chemotrophically grown cells of Rhodospirillum rubrum. On the basis of electron-microscopic investigations, chemical analysis, distribution of components involved in respiration, and poly-acrylamide gel electrophoresis, the heavy fraction (rho20 = 1.246 g per cm3) was identified as cell-wall, and the light fraction (rho = 1.145 g per cm3) as cyto-plasmic-membrane fragments. Electron micrographs showed cell-wall fragments as open structures while cytoplasmic-membrane preparations were composed of closed membrane vesicles. With respect to the main classes of chemical compounds, cell wall could be distinguished from cytoplasmic membranes by a rather low ratio of phospholipids per protein and a high ratio of carbohydrates per protein. The relative proportion of individual neutral sugars as well as phospholipids (except for lysophosphatidyl ethanolamine) revealed no significant differences between both envelope fractions. Fatty acid analysis demonstrated a higher proportion of saturated fatty acids in cell-wall than in cyto-plasmic-membrane fractions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the fractions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the fractions showed distinct protein compositions. While in cell-wall preparations polypeptides of 43,000 and 14,000 daltons predominated, 56,000- and 52,000-dalton polypeptides were the main protein subunits of cytoplasmic membranes. Cross contaminations of both cell-envelope fractions were defined.

Cell Fractionation↗
PubMed2002

Texture of cooked potatoes (Solanum tuberosum). 2. Changes in pectin composition during storage of potatoes.

During the storage season changes in the chemical composition of the pectin moiety of the cell walls of raw potatoes were studied. This compositional analysis was performed for the cultivars Nicola and Irene, which represent two extremes with regard to sensory-perceived texture. Both cultivars were divided into three size categories. From each size category a dry matter (DM) distribution was made. From these distributions potatoes at the low and high ends of this distribution were selected for further analysis. In total 12 different samples were analyzed three times during the storage season. The analysis comprised a pectin fractionation study. Pectic fractions were extracted from the cell wall material (CWM) by increasing the harshness of the extraction procedure. This resulted in a calcium-complexed pectic fraction, two pectic fractions weakly bound to the CWM, and a residue fraction, respectively. It was shown that no statistically significant differences (p > or = 0.95), either in yield or in chemical composition, could be observed between the two cultivars studied (Nicola and Irene), between sizes (large, medium, and small), and between potatoes with either high or low DM contents. However, statistically significant effects of storage both on the yield and on the chemical composition of the pectic moiety of the CWM could be observed, irrespective of cultivar, size, and DM content. Despite the substantial changes in the composition of the pectic moiety of the CWM of the raw material, no to minimal changes in the sensory-perceived texture of the cooked potatoes were observed upon storage. This suggests that the observed changes in pectin composition upon storage are overruled by other aspects that contribute more importantly to the sensory-perceived texture of steam-cooked potatoes.

Cell Wall↗
PubMed2005

Characterization of cell wall polysaccharides from the medicinal plant Panax notoginseng.

Panax notoginseng is a commonly used medicinal plant in south-western China. Recent studies indicate that wall polysaccharides are responsible for some of the immunostimulatory activity. Fractionation of the P. notoginseng root powder alcohol insoluble residue (AIR) and its compositional analysis enabled us to deduce the polysaccharide and protein composition of the root cell walls. P. notoginseng walls are composed primarily of polysaccharide (approximately 97% w/w) and some protein. The polysaccharides include pectic polysaccharides (neutral Type I 4-galactan (21%), arabinan (5%), acidic rhamnogalacturonan I (RG I, 2%) and homogalacturonan (HGA, 24%), non-cellulosic polysaccharides (heteroxylan, 3%), xyloglucan (XG, 3%) and heteromannan (1%)) and cellulose (24%). The root AIR also contains Type II AG/AGPs (5% w/w) typically associated with the plasma membrane and extracellular matrix. Thus, P. notoginseng roots contain polysaccharides typical of Type I primary cell walls but are distinguished by their very high levels of Type I 4-galactans and low levels of XGs. The major amino acids in the AIR were Leu (14 mol%), Asx (16 mol%), Glx (10 mol%), Ala (9 mol%), Thr (9 mol%) and Val (9 mol%).

Cell Wall↗
PubMed1999

In vitro synthesis and properties of pectin/Acetobacter xylinus cellulose composites

Pectin and cellulose are major components of most primary cell walls, yet little is known about the way in which they interact either during assembly or in subsequent functional performance of the wall. As a mimic of cell wall assembly, we studied the formation of molecular composites formed by deposition of cellulose from Acetobacter xylinus into pectin/calcium systems, and the molecular, architectural and mechanical properties of the composites obtained. The formation of interpenetrating cellulose/pectin composite networks (as envisaged in current models for primary cell walls) required a pre-existing, but not too strong, pectin network. For pectin either in solution or strongly networked, phase separation from cellulose occurred, providing two physical models for the formation of middle lamellae. Composite networks showed no evidence of direct molecular interaction between the components, but pectin networks became more aggregated following deposition of cellulose into them. The shear strength under small deformation conditions for cellulose/pectin composites was very similar to that of cellulose alone. In contrast, under uniaxial tension, extensibility was greatly increased and stiffness decreased. These major changes were due to the effect of pectin on cellulose network architecture at deposition, as they were maintained upon removal of the pectin component. These results show that the presence and physical state of pectin at the time of cellulose deposition in muro may be a significant determinant of subsequent extensibility without compromising strength.

Journal Article↗
PubMed2006

Investigation of toroidal pore and oligomerization by melittin using transmission electron microscopy.

We studied the effects of melittin on various cell wall components and vesicles of various lipid compositions. To interact with the cytoplasmic membrane, melittin must traverse the cell wall, which is composed of oligosaccharides. Here, we found that melittin had a strong affinity for chitin, peptidoglycan, and lipopolysaccharide. We further examined the influence of lipid compositions on the lysis of the membranes by melittin. The result showed that melittin bound better to negatively charged than to zwitterionic lipid vesicles but was more potent at inducing leakage from zwitterionic lipid vesicles. Our studies further indicated that the oligomeric state of melittin varied between tetramers and octamers during the formation of toroidal pores. Dextran leakage experiments confirmed the formation and dimension of these toroidal pores. Finally, transmission electron microscopy revealed that melittin formed pores via peptide oligomerization by the toroidal pore-forming mechanism. The toroidal pores composed of 7-8 nm diameter rings that encircled 3.5-4.5 nm diameter cavities on zwitterionic lipid vesicles.

Cell Membrane↗
PubMed2005

High-level (beta)-lactam resistance and cell wall synthesis catalyzed by the mecA homologue of Staphylococcus sciuri introduced into Staphylococcus aureus.

A close homologue of mecA, the determinant of broad-spectrum beta-lactam resistance in Staphylococcus aureus was recently identified as a native gene in the animal commensal species Staphylococcus sciuri. Introduction of the mecA homologue from a methicillin-resistant strain of S. sciuri into a susceptible strain of S. aureus caused an increase in drug resistance and allowed continued growth and cell wall synthesis of the bacteria in the presence of high concentrations of antibiotic. We determined the muropeptide composition of the S. sciuri cell wall by using a combination of high-performance liquid chromatography, mass spectrometric analysis, and Edman degradation. Several major differences between the cell walls of S. aureus and S. sciuri were noted. The pentapeptide branches in S. sciuri were composed of one alanine and four glycine residues in contrast to the pentaglycine units in S. aureus. The S. sciuri wall but not the wall of S. aureus contained tri- and tetrapeptide units, suggesting the presence of dd- and ld-carboxypeptidase activity. Most interestingly, S. aureus carrying the S. sciuri mecA and growing in methicillin-containing medium produced a cell wall typical of S. aureus and not S. sciuri, in spite of the fact that wall synthesis under these conditions had an absolute dependence on the heterologous S. sciuri gene product. The protein product of the S. sciuri mecA can efficiently participate in cell wall biosynthesis and build a cell wall using the cell wall precursors characteristic of the S. aureus host.

Anti-Bacterial Agents↗
PubMed2003

Feedback from the wall.

The ability of cells to perceive changes in the composition and mechanical properties of their cell wall is crucial for plants to achieve coordinated growth and development. Evidence is accumulating to show that the plant cell wall, like its yeast counterpart, is capable of triggering multiple signalling pathways. The components of the cell wall that are responsible for initiating these signal responses remain unknown; however, recent technological advances in cell wall analysis may now facilitate the identification of these components and accelerate the characterisation of changes that occur in cell wall mutants.

Arabidopsis Proteins↗
PubMed2006

Composition and cellular localization of tannins in Cabernet Sauvignon skins during growth.

For two successive years, cell walls were isolated from the internal part of skin cells of Vitis vinifera L. cv. Cabernet Sauvignon grape berries grown in a vineyard. Procyanidin localization and composition were determined over the course of development. Tannins were mainly localized in the inner cell fraction, due to their biosynthesis and storage. Cell wall tannins always exhibited a higher mean degree of polymerization as compared to the internal cell fraction, which had a constant mDP. The mDP of cell wall tannins also tended to increase at the end of maturation. Our results suggest tannin polymerization near the cell wall but an aggregation in the vacuole during growth. The tannin composition was typical of skins, and small differences were noted between the two cell parts. Surprisingly, epigallocatechin-3-gallate was also detected, although in a very small amount. Epicatechin was present in significant proportions in both fractions, especially as an extension subunit, while epigallocatechin was likewise abundant, also as a terminal subunit. Last, procyanidin composition and organization seemed to be characteristic of the Cabernet Sauvignon variety.

Biflavonoids↗
PubMed1993

Characterization of Candida albicans cell wall antigens with monoclonal antibodies.

The antigenic composition of Candida albicans is very complex. In order to study the antigenic relationship between blastoconidia and germ tubes of C. albicans, we produced several monoclonal antibodies and analyzed their reactivity against cell wall antigens either in intact cells or in cells treated with dithiothreitol. Overall, four types of reactivity were found. Monoclonal antibodies 3D9 and 15C9 stained the germ tubes only when tested by indirect immunofluorescence. However, they showed a different reactivity by immunoblotting. Monoclonal antibody 3D9 reacted with antigens with molecular masses of > 200 and 180 kDa specifically expressed in the germ tube. Monoclonal antibody 15C9 reacted with antigens of 87, 50, and 34 kDa present in the germ tube extract and with antigens of 92, 50, 34, and 32 kDa present in the blastoconidium extract. The reactivity of blastoconidia treated for different times with dithiothreitol with these monoclonal antibodies was also studied by enzyme-linked immunosorbent assay. The reactivity of monoclonal antibody 3D9 did not significantly change during the cell wall extraction. However, the reactivity of monoclonal antibody 15C9 was increased for blastoconidia extracted for 60 min and decreased markedly for blastocondia extracted for 120 min. Monoclonal antibody G3B was nonreactive by indirect immunofluoresence but reacted with antigens of 47 and 38 kDa present in the germ tube extract and with an antigen of 47 kDa present in the blastoconidium extract. Monoclonal antibody B9E stained both morphological phases by indirect immunofluorescence. By immunoblotting, it reacted with antigens of > 70 kDa present in the germ tube extract and with antigens of > 63, 56, 47, and 38 kDa present in the blastoconidium extract. Based on the results presented in this study, four types of antigens are described. Type I antigens are expressed on the outermost layers of the germ tube cell wall only. Type II antigens are expressed both on the germ tube cell wall surface and within the blastoconidium cell wall. Type III antigens are found within the cell wall of both blastoconidia and germ tubes. Type IV antigens are expressed on both the blastoconidium and germ tube surface. Two types more can be hypothesized for antigens expressed on the blastoconidium cell surface and within the germ tube cell wall (type V) and for those expressed on the blastoconidium surface only (type VI).

Animals↗
PubMed2006

The Populus homeobox gene ARBORKNOX1 reveals overlapping mechanisms regulating the shoot apical meristem and the vascular cambium.

Secondary growth is supported by a dividing population of meristematic cells within the vascular cambium whose daughter cells are recruited to differentiate within secondary phloem and xylem tissues. We cloned a Populus Class 1 KNOX homeobox gene, ARBORKNOX1 (ARK1), which is orthologous to Arabidopsis SHOOT MERISTEMLESS (STM). ARK1 is expressed in the shoot apical meristem (SAM) and the vascular cambium, and is down-regulated in the terminally differentiated cells of leaves and secondary vascular tissues that are derived from these meristems. Transformation of Populus with either ARK1 or STM over-expression constructs results in similar morphological phenotypes characterized by inhibition of the differentiation of leaves, internode elongation, and secondary vascular cell types in stems. Microarray analysis showed that 41% of genes up-regulated in the stems of ARK1 over-expressing plants encode proteins involved in extracellular matrix synthesis or modification, including proteins involved in cell identity and signaling, cell adhesion, or cell differentiation. These gene expression differences are reflected in alterations of cell wall biochemistry and lignin composition in ARK1 over-expressing plants. Our results suggest that ARK1 has a complex mode of action that may include regulating cell fates through modification of the extracellular matrix. Our findings support the hypothesis that the SAM and vascular cambium are regulated by overlapping genetic programs.

Amino Acid Sequence↗
PubMed1969

Specificity of a bacteriolytic enzyme from Pseudomonas aeruginosa.

A bacteriolytic enzyme isolated from shake-flask cultures of Pseudomonas aeruginosa and capable of lysing cells of Staphylococcus aureus was purified approximately 500-fold by passage through diethylaminoethyl cellulose and chromatography on carboxymethyl-cellulose. The purified enzyme was shown to act as an endopeptidase, cleaving the pentaglycine cross-bridges of the cell wall peptidoglycan at d-alanyl-glycine and glycyl-glycine linkages with the release of di-, tri-, and tetraglycine fragments. Release of NH(2)-alanine indicated weak N-acetylmuramyl-l-alanine amidase activity, but most of the residual peptide remained attached to the glycan. No hydrolysis of the glycan occurred. The lytic spectrum of the enzyme toward a variety of other cell walls of known peptidoglycan composition indicated relatively high specificity for peptidoglycans with polyglycine bridges.

Aminohydrolases↗
PubMed1989

[The effect of blood serum components on the growth, biochemical and biological properties of streptococcus].

The components of cattle blood serum, added to the medium for the cultivation of group A streptococci, considerably decrease the period of adaptation and increase the balanced growth rate of streptococci, which is manifested by changes in the surface structures of the cell wall: the absence or modification of protein M. Streptococci grown under these conditions lose their capacity for phagocytosis, and from the cell walls obtained from these streptococci no surface protein M can be isolated by pepsin treatment. Nevertheless, the ratio of the main cell-wall components (proteins, polysaccharide and peptidoglycan), the amino acid composition, as well as the resistance of the cell walls to the action of trypsin and endo-N-acetylmuramidase are the same in M+ and Mx variants, that makes it possible to infer that the modification of protein M or the inhibition of its synthesis occurs during the growth of streptococci in the presence of blood serum components.

Amino Acids↗
PubMed1996

STRUCTURE AND BIOGENESIS OF THE CELL WALLS OF GRASSES.

The chemical structures of the primary cell walls of the grasses and their progenitors differ from those of all other flowering plant species. They vary in the complex glycans that interlace and cross-link the cellulose microfibrils to form a strong framework, in the nature of the gel matrix surrounding this framework, and in the types of aromatic substances and structural proteins that covalently cross-link the primary and secondary walls and lock cells into shape. This review focuses on the chemistry of the unique polysaccharides, aromatic substances, and proteins of the grasses and how these structural elements are synthesized and assembled into dynamic and functional cell walls. Despite wide differences in wall composition, the developmental physiology of grasses is similar to that of all flowering plants. Grass cells respond similarly to environmental cues and growth regulators, exhibit the same alterations in physical properties of the wall to allow cell growth, and possess similar patterns of wall biogenesis during the development of specific cell and tissue types. Possible unifying mechanisms of growth are suggested to explain how grasses perform the same wall functions as other plants but with different constituents and architecture.

Journal Article↗
PubMed1999

Responses of Mentha suspension-cultured cells to 2,4-dichlorophenoxyacetic acid and accumulation of esterified phenolic acids in their cell walls.

Two distinct types of cell growth of suspension-cultured Mentha were formed when the cells maintained in the medium containing 1000 micrograms l-1 2,4-D were subcultured into different 2,4-D concentrations. Few cell elongation of Mentha (average cell length: 34-40 microns) was observed after division in the medium containing 1-200 micrograms l-1 2,4-D; and significant cell elongation (average cell length: 95-130 microns) was observed after cell division in the medium containing 500-2000 micrograms l-1 2,4-D. A close correlation between culture medium and water content in the cells indicated that 2,4-D promoted cell elongation by water uptake. Amounts of phenolic acid in cell walls were much higher in unelongated cell walls than in elongated ones during the cultivation, and there was a close correlation between the amounts and the level of PAL activity in elongated and unelongated cells. However, there was no significant difference in cell wall components and its neutral sugar composition between elongated and unelongated cells.

2,4-Dichlorophenoxyacetic Acid↗
PubMed2005

Differential expression of chitin synthase (CHS) and glucan synthase (FKS) genes correlates with the formation of a modified, thinner cell wall in in vivo-produced Beauveria bassiana cells.

During infection (in vivo), the entomopathogenic fungus Beauveria bassiana produces yeast-like cells that are surrounded by modified cell walls. These modifications have been related to the fungus ability to limit recognition by the host defense system. The composition of the in vivo cell wall was analyzed using a combination of cytochemical and molecular techniques. The in vivo cell walls still contained both chitin and 1,3-beta-glucan, but they were significantly thinner than in vitro cell walls (50-60 nm versus 100-160 nm, respectively). The difference in cell wall thickness was correlated with transcriptional regulation of cell wall-related genes: quantitative RT-PCR reactions demonstrated that B. bassiana chitin synthase (CHS) and glucan synthase (FKS) genes are down regulated in vivo. These analyses indicate that in vivo-triggered phenotypic modifications, including cell wall adjustments, are controlled by molecular mechanisms that include regulation of gene expression at the transcriptional level.

Amino Acid Sequence↗