-
The Journal of Biological Chemistry May 1970
Comparative Study
Topics: Amino Acids; Chemical Phenomena; Chemistry; Chromatography, DEAE-Cellulose; Chromatography, Gel; Coenzymes; Electrophoresis; Hot Temperature; Hydrogen-Ion Concentration; Kinetics; Methods; Molecular Weight; Oxidoreductases; Peptides; Proteins; Saccharomyces; Spectrophotometry; Streptomycin; Sulfides; Ultracentrifugation; Ultraviolet Rays
PubMed: 5442277
DOI: No ID Found -
The Journal of Biological Chemistry Jan 1977Raw extract in 2 m CaCl2 of bovine nasal septum cartilage was eluted from 4 per cent agarose gel to give a "void volume" Fraction v-4, which was indistinguishable in...
Raw extract in 2 m CaCl2 of bovine nasal septum cartilage was eluted from 4 per cent agarose gel to give a "void volume" Fraction v-4, which was indistinguishable in composition and behavior on viscometric and sedimentation analysis from the densest fraction obtained by associative centrifugation in a cesium chloride density gradient. The sulfated proteoglycan was precipitated (Fraction A) by cetylpyridinium chloride from acidic solutions of Fraction v-4 or of dialyzed raw ectract. Neutralization under conditions of low ionic strength precipitated a further small fraction (B), which contained from 0.5 to 1 per cent of the uronic acid in the original extract. Analysis by associative and dissociative density gradient centrifugation demonstrated that Fraction B resembled in effective density known samples of hyaluronic acid from other sources. Gel chromatography of proteolytic digests of Fractions A and B on 6 per cent agarose indicated that cetylpyridinium chloride precipitation essentially separated sulfated proteoglycan (A) from hyaluronic acid (B). A viscosity-average molecular weight of about 5 x 10(5) was estimated for a sample of Fraction B purified in a dissociative (4 M guanidine hydrochloride + CsCl) density gradient. Sedimentation velocity data were consistent with this result. Analysis of hexosamines showed that the sample contained 96 per cent glucosamine, confirming the identification of hyaluronic acid. The proteoglycan fraction (A) resembled "subunits" in its sedimentation behavior.
Topics: Amino Acids; Animals; Cartilage; Cattle; Cetylpyridinium; Chemical Precipitation; Hexosamines; Hexoses; Hyaluronic Acid; Methods; Nasal Septum; Proteins; Uronic Acids
PubMed: 833135
DOI: No ID Found -
Journal of Bacteriology Feb 1976A new heteropolysaccharide has been isolated by alkaline extraction of hyphal walls of Aspergillus niger NRRL 326 grown in surface culture. Its composition by weight, as...
A new heteropolysaccharide has been isolated by alkaline extraction of hyphal walls of Aspergillus niger NRRL 326 grown in surface culture. Its composition by weight, as determined by paper and gas chromatography and colorimetric analyses, is 70% galactose, 20% galactosamine, 6% glucose, and 1% acetyl. Two independent experiments have been used to ascertain copolymer structure: permeation chromatography in 6 M guanidinium hydrochloride, with controlled-pore glass columns of two fractionation ranges, and nitrous acid deaminative cleavage of galactosaminogalactan followed by reduction of fragments with [3H]borohydride and gel filtration chromatography. One of the tritiated fragments is tentatively identified as the disaccharide derivative galactopyranosyl 2,5-anhydrotalitol, on the basis of chromatographic properties and by kinetics of its acid hydrolysis. Smith degradation, methylation, deamination, and optical rotation studies indicate that the galactosaminogalactan consists of a linear array of hexopyranosyl units joined almost exclusively by alpha-(1 leads to 4) linkages. Hexosaminyl moieties are distributed randomly along the chains, which have an average degree of polymerization of about 100. The possible significance of this macromolecule in hyphal structure is considered.
Topics: Aspergillus; Aspergillus niger; Cell Wall; Chemical Precipitation; Chromatography; Galactosamine; Galactose; Glucose; Glycosides; Hydrolysis; Methylation; Oxidation-Reduction; Periodic Acid; Polysaccharides
PubMed: 173713
DOI: 10.1128/jb.125.2.655-669.1976 -
The Biochemical Journal Apr 19751. New preparations of reduced carboxymethylated beta-ovomucin (S-carboxymethyl-beta-ovomucin) were homogeneous by sedimentation analysis, analytical sedimentation to...
1. New preparations of reduced carboxymethylated beta-ovomucin (S-carboxymethyl-beta-ovomucin) were homogeneous by sedimentation analysis, analytical sedimentation to equilibrium in CsCl gradients, and disc electrophoresis in sodium dodecyl sulphate. 2. Degradation of S-carboxymethyl-beta-ovomucin with either CNBr or trypsin indicated the presence of a subunit (approx. mol. wt. 112300). 3. Electron microscopy showed that S-carboxymethyl-beta-ovomucin consisted of chains of globular units (approx. mol. wt. 103 000). IN 6M-guanidinium chloride S-carboxymethyl-beta-ovomucin existed mainly as an aggregate (mol. wt. 720 000). 4. S-Carboxymethyl-beta-ovomucin contained ester sulphate (4.24%, W/W) and carbohydrate (60%, W/W), which consisted of large amounts of galactose (22%, W/W), galactosamine (8.9%, W/W) and sialic acid (10.6%, W/W). 5. An unreduced soluble fibrous component (component SGH) extracted from crude ovomucin precipitate with 5M-guanidinium chloride contained beta-ovomucin (approx. 70%, W/W). By using the Scheraga-Mandelkern equation the molecular weight of component SGH was calculated to be 11.5 times 10(6).
Topics: Amino Acids; Animals; Carbohydrates; Carboxylic Acids; Centrifugation; Chickens; Cyanogen Bromide; Egg Proteins; Electrophoresis, Disc; Esters; Galactosamine; Galactose; Microscopy, Electron; Molecular Weight; Ovalbumin; Ovomucin; Oxidation-Reduction; Sialic Acids; Trypsin; Viscosity
PubMed: 1171686
DOI: 10.1042/bj1470055 -
FEBS Letters Oct 1972
Topics: Amino Acid Sequence; Amino Acids; Animals; Carboxypeptidases; Chromatography, Gas; Crystallization; Ducks; Electrophoresis, Disc; Glucagon; Pancreas; Thiocyanates
PubMed: 4636745
DOI: 10.1016/0014-5793(72)80595-7 -
Journal of Lipid Research Mar 1971The quantitative isolation of total glycosphingolipids from crude lipid extracts without contamination from other lipid classes is described. The method consists of (a)... (Comparative Study)
Comparative Study
The quantitative isolation of total glycosphingolipids from crude lipid extracts without contamination from other lipid classes is described. The method consists of (a) acetylation of total lipids with pyridine and acetic anhydride, (b) separation of acetylated glycolipids from nonglycolipids on a magnesia-silica gel (Florisil) column, and (c) deacetylation of glycolipid in chloroform-methanol-sodium methoxide. This method is useful for determination of microgram quantities of glycolipids derived from less than 1 ml of packed cells.
Topics: Acetates; Acetone; Anhydrides; Animals; Carcinoma, Hepatocellular; Cell Biology; Cerebrosides; Chloroform; Chromatography; Fibroblasts; Glycolipids; Hydrocarbons, Halogenated; Kidney; Lipids; Liver; Liver Neoplasms; Methanol; Methods; Microchemistry; Phospholipids; Pyridines; Rats; Sphingolipids; Spleen
PubMed: 4324310
DOI: No ID Found -
The Journal of Biological Chemistry Nov 1973
Topics: Amino Acid Sequence; Amino Acids; Animals; Chemical Phenomena; Chemistry; Chromatography, Ion Exchange; Cysteine; Disulfides; Electrophoresis, Paper; Epithelium; Growth Substances; Male; Mice; Peptide Fragments; Peptides; Skin; Solubility; Submandibular Gland; Thermolysin
PubMed: 4750422
DOI: No ID Found -
The Journal of Biological Chemistry Jan 1981
Topics: Amino Acids; Animals; C-Reactive Protein; Carbohydrates; Chromatography, High Pressure Liquid; Electron Spin Resonance Spectroscopy; Hemagglutination Tests; Hemolymph; Horseshoe Crabs; Immunodiffusion; Lectins; Molecular Weight; Spin Labels
PubMed: 6256375
DOI: No ID Found -
The Biochemical Journal Sep 19741. The mucoprotein from pig gastric mucus has been purified by equilibrium centrifugation in a CsCl gradient. 2. This procedure removes the non-covalently bound protein,...
1. The mucoprotein from pig gastric mucus has been purified by equilibrium centrifugation in a CsCl gradient. 2. This procedure removes the non-covalently bound protein, which is closely associated with the mucoprotein and not easily removed from it by gel filtration. 3. The purified mucoprotein is separable by gel filtration into a high-molecular-weight mucoprotein A (mol.wt. 2.3x10(6)) and a low-molecular-weight mucoprotein B/C (mol.wt. 1.15x10(6)). 4. These two mucoproteins have the same chemical analysis namely fucose 11.3%, galactose 26%, glucosamine 19.5%, galactosamine 8.3% and protein 13.6%. 5. Mucoprotein A contains 3.1% ester sulphate. 6. These mucoproteins are isolated without enzymic digestion and have a higher protein content than the blood-group-substance mucoproteins from proteolytic digestion of gastric mucus. Detailed amino acid analysis shows that the extra protein in the non-enzymically digested material is composed of amino acids other than serine and threonine. 7. Mucoproteins A and B/C contain respectively 130 and 9 half-cystine residues per molecule of which about 78 and 6 residues are involved in disulphide linkages. 8. Cleavage of these disulphide linkages by mercaptoethanol splits both mucoproteins into four equally sized subunits of mol.wt. 5.2x10(5) for mucoprotein A and 2.8x10(4) for mucoprotein B/C. 9. The sole N-terminal amino acid of mucoprotein A is aspartic acid, whereas mucoprotein B/C has several different N-terminal amino acid residues.
Topics: Amino Acids; Animals; Aspartic Acid; Centrifugation, Density Gradient; Cystine; Disulfides; Esters; Fucose; Galactosamine; Galactose; Gastric Mucosa; Glucosamine; Mercaptoethanol; Molecular Weight; Mucoproteins; Proteins; Sulfates; Swine
PubMed: 4463953
DOI: 10.1042/bj1410633 -
European Journal of Biochemistry May 1976From Escherichia coli 0124 two lipopolysaccharide preparations were obtained with phenol/water extraction and cetavlon precipitation. Polyacrylamide gel electrophoresis...
From Escherichia coli 0124 two lipopolysaccharide preparations were obtained with phenol/water extraction and cetavlon precipitation. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate and chemical analysis showed that the two preparations from E. coli 0124 and the corresponding preparations from Shigella dysenteriae type 3 reacted alike. The O-specific polysaccharide moiety was characterized with proton magnetic resonance spectroscopy, optical rotation and paper electrophoresis. The constituents were determined by gas chromatography and ion-exchange chromatography. The polysaccharide contained glucose (Glc), galactose (Gal), galactosamine (GalN) and 4-O-(1'-carboxyethyl)-D-glucopyranose (glucolactilic acid, GlcLA) in the molar ratios of 1:2:1:1. Glucolactilic acid, which has a structure similar to muramic acid, was first found in Sh. dysenteriae. The polysaccharide from E. coli 0124 and oligosaccharides obtained from it by partial acid hydrolysis were subjected to methylation analysis using the method of combined gas chromatography--mass spectrometry. The results indicated that the pentasaccharide repeating unit of the polysaccharide is (see article). In the polysaccharide the repeating units are joined through galactofuranosidic linkages. This structure is identical with that of the somatic polysaccharide of Sh. dysenterae type 3.
Topics: Cell Wall; Escherichia coli; Galactosamine; Galactose; Glucose; Hemagglutination Tests; Immunodiffusion; Lipopolysaccharides; Molecular Conformation; Oligosaccharides; Polysaccharides, Bacterial; Shigella; Sugar Acids
PubMed: 819266
DOI: 10.1111/j.1432-1033.1976.tb10327.x