scholarly journals The structure of C-polysaccharide from the walls of Streptococcus pneumoniae

1978 ◽  
Vol 175 (3) ◽  
pp. 1033-1042 ◽  
Author(s):  
I R Poxton ◽  
E Tarelli ◽  
J Baddiley
Keyword(s):  
Cell Walls ◽  
Active Component ◽  
Alkali Treatment ◽  
Teichoic Acid ◽  
Periodate Oxidation ◽  
Structural Features ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Chemical Degradation ◽  
Acid And Alkali Treatment

The well-known immologically active component of pneumococci, C-polysaccharide, is a teichoic acid that can be isolated from the cell walls and purified by Sephadex and ion-exchange chromatography. Further details of the structure of C-teichoic acid were established by chemical degradation, including hydrolysis in acid and alkali, treatment with HF, periodate oxidation and methylation. In addition, the use of 13C n.m.r. has confirmed some of these structural features and resulted in a proposal for the order of substituents, the location of positions of substitution and the configuration of anomeric centres in the repeating unit of the polymer.

scholarly journals STRUCTURAL AND CHEMICAL CHARACTERISTIC OF PECTIN FROM PICEA ABIES GREENERY

2020 ◽  
pp. 59-71
Author(s):  
Evgeniy Gennad'yevich Shakhmatov ◽  
Elena Nikolayevna Makarova
Keyword(s):  
Cell Walls ◽  
New Technologies ◽  
Chemical Characteristic ◽  
Structural Features ◽  
Exchange Chromatography ◽  
Raw Material ◽  
Rhamnogalacturonan I ◽  
Wood Processing ◽  
Potential Source ◽  
Pectin Substances

The present work aimed to determine structural features of polysaccharides derived from the P. abies foliage by extraction with a (NH4)2C2O4 solution. The isolated polysaccharide was studied in detail by the methods of ion exchange chromatography, partial acidic hydrolys and NMR spectroscopy. It was shown that this polysaccharide contained polymers of various structures. The major constituents of PAO were low-methoxyl and low-acetylated 1,4-a-D-galacturonan and by minor parts of partly 2-O- and/or 3-O- acetylated rhamnogalacturonan-I (RG-I). The side carbohydrate chains of the branched region of RG-I were represented predominantly by highly branched 1,5-a-L-arabinan and minor portions of 1,4-β-D-galactan. In addition to the dominant pectins, polysaccharide PAO contained binding glycans of the glucomannans class, which indicated a close interaction of these polysaccharides in the cell walls. Thus, the structural features of pectin woody P. abies, extracted with a solution of (NH4)2C2O4, were first determined. It can be concluded that P. abies woody greens, a large tonnage waste from the wood processing industry, can be considered as a potential source of pectin substances. The results of studying the structure of components of woody green P. abies can be the basis for the development and improvement of new technologies for the integrated use of this raw material.

scholarly journals Cell-wall polysaccharides and glycoproteins of parenchymatous tissues of runner bean (Phaseolus coccineus)

1990 ◽  
Vol 269 (2) ◽  
pp. 393-402 ◽  
Author(s):  
P Ryden ◽  
R R Selvendran
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Anion Exchange Chromatography ◽  
Structural Features ◽  
Good Evidence ◽  
Exchange Chromatography ◽  
Cell Wall Polysaccharides ◽  
Pectic Polysaccharides ◽  
Runner Bean ◽  
Cellulose Residue

1. Polymers were solubilized from the cell walls of parenchyma from mature runner-bean pods with minimum degradation by successive extractions with cyclohexane-trans-1,2-diamine-NNN′N′-tetra-acetate (CDTA), Na2CO3 and KOH to leave the alpha-cellulose residue, which contained cross-linked pectic polysaccharides and Hyp-rich glycoproteins. These were solubilized with chlorite/acetic acid and cellulase. The polymers were fractionated by anion-exchange chromatography, and fractions were subjected to methylation analysis. 2. The pectic polysaccharides differed in their ease of extraction, and a small proportion were highly cross-linked. The bulk of the pectic polysaccharides solubilized by CDTA and Na2CO3 were less branched than those solubilized by KOH. There was good evidence that most of the pectic polysaccharides were not degraded during extraction. 3. The protein-containing fractions included Hyp-rich and Hyp-poor glycoproteins associated with easily extractable pectic polysaccharides, Hyp-rich glycoproteins solubilized with 4M-KOH+borate, the bulk of which were not associated with pectic polysaccharides, and highly cross-linked Hyp-rich glycoproteins. 4. Isodityrosine was not detected, suggesting that it does not have a (major) cross-linking role in these walls. Instead, it is suggested that phenolics, presumably linked to C-5 of 3,5-linked Araf residues of Hyp-rich glycoproteins, serve to cross-link some of the polymers. 5. There were two main types of xyloglucan, with different degrees of branching. The bulk of the less branched xyloglucans were solubilized by more-concentrated alkali. The anomeric configurations of the sugars in one of the highly branched xyloglucans were determined by 13C-n.m.r. spectroscopy. 6. The structural features of the cell-wall polymers and complexes are discussed in relation to the structure of the cell walls of parenchyma tissues.

scholarly journals Co-polymeric glycosaminoglycans in transformed cells. Transformation-dependent changes in the co-polymeric structure of heparan sulphate

1982 ◽  
Vol 201 (1) ◽  
pp. 233-240 ◽  
Author(s):  
Lars-Ȧke Fransson ◽  
Birgitta Havsmark ◽  
Vincenzo P. Chiarugi
Keyword(s):  
Heparan Sulphate ◽  
Periodate Oxidation ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Transformed Cells ◽  
Large Contribution ◽  
3T3 Cells ◽  
Normal Case ◽  
3T3 Fibroblasts ◽  
Lower Charge

1. Heparan sulphates from normal 3T3 fibroblasts are association-prone as indicated by their affinity for agarose gels substituted with cognate heparan sulphate species. Heparan sulphates from SV40-transformed or polyoma-virus-transformed cells have no affinity for the same gels. 2. Heparan sulphates from the medium, the pericellular and intracellular pools of normal, SV40-transformed and polyoma-transformed 3T3 cells were separated into four subfractions (HS1–HS4) by ion-exchange chromatography. In general, HS1–HS3 were found in cell-derived heparan sulphates, whereas HS3–HS4 were present in the medium. The heparan sulphates from transformed cells were more heterogeneous and of lower charge density than those from the normal counterpart. 3. Degradations via periodate oxidation/alkaline elimination yielded the oligomers glucosamine-(hexuronate–glucosamine)n-R with n=1–5 and a large proportion of N-sulphate groups. There was a large contribution of fragments n=4–5 from heparan sulphates of normal cells. These fragments were less common in low-sulphated heparan sulphates of transformed cells. In the case of medium-drived heparan sulphates all species had a low content of fragments n=4–5. 4. The size distribution of (glucuronate–N-acetylglucosamine)n regions was assessed after deaminative cleavage. It was broad and ranged from n=1–10 for all heparan sulphate species. In the case of medium-derived heparan sulphates there were distinct differences between normal and transformed cells. In the latter chains the N-acetyl-rich segments were both shorter and longer than in the normal case. The shape of the disaccharide peak was consistent with a lower content of O-sulphate in the heparan sulphates from transformed cells. 5. It was concluded that heparan sulphates from medium or transformed cells exhibit the greatest structural deviation from the normal case. The finding of lower proportions of extended, iduronate/glucuronate-bearing, N-sulphate-rich segments in heparan sulphates of transformed cells was particularly interesting in view of the fact that these elements have been associated with ability to self-interact.

scholarly journals The presence of ribonucleic acid in the cell walls of higher plants

1968 ◽  
Vol 108 (1) ◽  
pp. 25-31 ◽  
Author(s):  
P. D. Phethean ◽  
L. Jervis ◽  
Mary Hallaway
Keyword(s):  
Cell Wall ◽  
Ion Exchange ◽  
Ribosomal Rna ◽  
Cell Walls ◽  
Potassium Hydroxide ◽  
Base Composition ◽  
Higher Plants ◽  
Nuclear Material ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography

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° 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.

scholarly journals The teichuronic acid from the walls of Bacillus licheniformis A.T.C.C. 9945

1980 ◽  
Vol 191 (2) ◽  
pp. 305-318 ◽  
Author(s):  
M R Lifely ◽  
E Tarelli ◽  
J Baddiley
Keyword(s):  
Ion Exchange ◽  
Bacillus Licheniformis ◽  
Cell Walls ◽  
Periodate Oxidation ◽  
Structural Features ◽  
Phosphate Limitation ◽  
Detailed Structure ◽  
Methylation Analysis ◽  
Partial Acid Hydrolysis ◽  
Teichuronic Acid

The teichuronic acid of Bacillus licheniformis A.T.C.C. 9945 grown under phosphate limitation was isolated from the cell walls and purified by ion-exchange and Sephadex chromatography. The detailed structure of the polysaccharide was established by methylation analysis, periodate oxidation and partial acid hydrolysis. The polymer is composed of tetrasaccharide repeating units with the structure [GlcA beta(1 leads to 4)GlcA beta(1 leads to 3)GalNAc beta(1 leads to 6)GalNAc alpha(1 leads to 4)n. 13C n.m.r. analysis has confirmed most of the structural features of the polysaccharide and, in particular, the anomeric configurations and linkage positions of substituents. The teichuronic acid from glucose-limited cells was identical with that from cells grown under phosphate limitation.

scholarly journals STUDIES ON THE IMMUNOCHEMISTRY OF STREPTOCOCCAL MUCOPEPTIDE

1966 ◽  
Vol 124 (2) ◽  
pp. 155-171 ◽  
Author(s):  
Walter W. Karakawa ◽  
Richard M. Krause
Keyword(s):  
Cell Walls ◽  
Antigenic Determinant ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Residues ◽  
Precipitin Reaction ◽  
Peptide Moiety ◽  
Streptomyces Albus ◽  
D Cell ◽  
Group D

Streptococcal mucopeptide, solubilized by either ultrasonic treatment or lysozyme, gave a precipitin reaction with rabbit antimucopeptide serum. A haptenic inhibitor of this reaction, which was composed of alanine, glutamic acid, and lysine in a mole ratio of 4:1:1, was isolated from a Streptomyces albus enzymes digest of Group D cell walls by ion exchange chromatography. When selected antisera were employed, greater than 90% inhibition of the mucopeptide quantitative precipitin reaction was achieved with 2 mg/ml of this inhibitor, whereas a hexosamine fraction with minimal concentrations of amino acid residues was inactive in this respect. These results suggest that the peptide moiety is an antigenic determinant of mucopeptide. Preliminary results indicate that the hexosamine polymer of the mucopeptide is a secondary antigenic determinant.

scholarly journals Ion-exchange purification and structural characterization of five sulfated fucoidans from brown algae

Glycobiology ◽  
2020 ◽  
Author(s):  
Andreas Sichert ◽  
Sophie Le Gall ◽  
Leesa Jane Klau ◽  
Brigitte Laillet ◽  
Hélène Rogniaux ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Brown Algae ◽  
Structural Diversity ◽  
Structural Features ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sulfated Polysaccharides ◽  
Monomer Composition ◽  
Novel Structure ◽  
Nmr Characterization

Abstract Fucoidans are a diverse class of sulfated polysaccharides integral to the cell wall of brown algae, and due to their various bioactivities, they are potential drugs. Standardized work with fucoidans is required for structure–function studies, but remains challenging since available fucoidan preparations are often contaminated with other algal compounds. Additionally, fucoidans are structurally diverse depending on species and season, urging the need for standardized purification protocols. Here, we use ion-exchange chromatography to purify different fucoidans and found a high structural diversity between fucoidans. Ion-exchange chromatography efficiently removes the polysaccharides alginate and laminarin and other contaminants such as proteins and phlorotannins across a broad range of fucoidans from major brown algal orders including Ectocarpales, Laminariales and Fucales. By monomer composition, linkage analysis and NMR characterization, we identified galacturonic acid, glucuronic acid and O-acetylation as new structural features of certain fucoidans and provided a novel structure of fucoidan from Durvillaea potatorum with α-1,3-linked fucose backbone and β-1,6 and β-1,3 galactose branches. This study emphasizes the use of standardized ion-exchange chromatography to obtain defined fucoidans for subsequent molecular studies.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

Preparation and Characterization of NH2-Terminal Fibrinogen Bβ Fragments from N-DSK of Human Fibrinogen

1984 ◽  
Vol 51 (01) ◽  
pp. 016-021 ◽  
Author(s):  
S Birken ◽  
G Agosto ◽  
B Lahiri ◽  
R Canfield
Keyword(s):  
High Performance ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Human Fibrinogen ◽  
Human Volunteers ◽  
Cnbr Cleavage ◽  
Two Peaks

SummaryIn order to investigate the early release of NH2-terminal plasmic fragments from the Bβ chain of fibrinogen, substantial quantities of Bβ 1-42 and Bβ 1-21 are required as immunogens, as radioimmunoassay standards and for infusion into human volunteers to determine the half-lives of these peptides. Towards this end methods that employ selective proteolytic cleavage of these fragments from fibrinogen have been developed. Both the N-DSK fragment, produced by CNBr cleavage of fibrinogen, and Bβ 1-118 were employed as substrates for plasmin with the finding of higher yields from N-DSK. Bβ 1-42 and Bβ 1-21 were purified by gel filtration and ion-exchange chromatography on SP-Sephadex using volatile buffers. When the purified preparation of Bβ 1-42 was chromatographed on reverse-phase high performance liquid chromatography, two peaks of identical amino acid composition were separated, presumably due either to pyroglutamate or to amide differences.

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