Showing papers on "Paper chromatography published in 1984"
TL;DR: It is concluded that a point mutation in the human ALDH2 locus produced the glutamine leads to lysine substitution and enzyme inactivation in atypical livers.
Abstract: Usual human livers contain two major aldehyde dehydrogenase [(ALDH) aldehyde:NAD+ oxidoreductase] isozymes--i.e., a cytosolic ALDH1 component and a mitochondrial ALDH2 component--whereas approximately equal to 50% of Orientals are "atypical" and have only the ALDH1 isozyme and are missing the ALDH2 isozyme. We previously demonstrated that atypical livers contain an enzymatically inactive but immunologically crossreactive material (CRM) corresponding to the ALDH2 component. The enzymatically active ALDH2 obtained from a usual liver and the CRM obtained from an atypical liver were reduced, S-carboxymethylated, and digested by trypsin. Separation of their digests by high-performance reverse-phase chromatography and by two-dimensional paper chromatography and electrophoresis revealed that ALDH2 contained a peptide sequence of -Glu-Leu-Gly-Glu-Ala-Gly-Leu-Gln-Ala-Asn-Val-Gln-Val-Lys- and that the glutamine adjacent to lysine was substituted by lysine in CRM. All other tryptic peptides, including eight peptides containing S-carboxymethylcysteine, were common in ALDH2 and CRM. It is concluded that a point mutation in the human ALDH2 locus produced the glutamine leads to lysine substitution and enzyme inactivation.
421 citations
TL;DR: A rapid ion-exchange method has been used to purify the alginate from the extracellular material of mucoid strains of Pseudomonas aeruginosa isolated from the lungs of cystic fibrosis patients, and the chemical/physical properties are considered in the context of the growth of the organism in the lung.
126 citations
76 citations
TL;DR: The results indicate that ferredoxin-linked glutamate synthase of spinach leaves is an iron-sulfur flavoprotein.
Abstract: Ferredoxin-dependent glutamate synthase (native enzyme) [EC 1.4.7.1] of spinach has been purified to homogeneity in the presence of 2-oxoglutarate and sodium chloride and the properties of the enzyme have been studied. The molecular weight of the enzyme was estimated to be 140,000 by gel filtration. Subunit analysis by SDS-gel electrophoresis yielded a single protein band whose molecular weight was about 170,000. This purified enzyme showed a flavo-protein-like absorption spectrum having maxima at 279 and 438 nm with shoulders at 415 and 460 nm and a broad band around 360 nm. Fluorometric data indicated the presence of 2 mol of flavin per mol of the enzyme. Preliminary paper chromatography results indicated the presence of FAD and FMN in the purified enzyme. The enzyme also contained 4 mol of acid-labile sulfide and 4 g-atoms iron per mol of enzyme. In the absence of 2-oxoglutarate and/or sodium chloride, the purified enzyme was separated by either DE-52 cellulose chromatography or gel filtration with Ultrogel AcA 34 into two molecular forms (modified enzymes) with considerable inactivation. When reduced methyl viologen plus ferredoxin was used as the electron donor, the purified (native) enzyme showed high ferredoxin-dependent activity with a specific activity of 100 units/mg protein. Methyl viologen-dependent activity was negligible in the absence of ferredoxin. Kinetic properties and results of ESR studies were described. The results indicate that ferredoxin-linked glutamate synthase of spinach leaves is an iron-sulfur flavoprotein.
43 citations
TL;DR: A branched oligosaccharide was prepared by limited action of Bacillus macerans cycloamylose glucanotransferase (BME) on waxy corn starch as mentioned in this paper.
41 citations
TL;DR: A patient with a lysosomal storage disease, progressive neurologic degeneration, and renal failure was found to have accumulated a low molecular weight ninhydrin and phenol-H2SO4 reactive compound, which is the linkage region in ADP-ribosylation of histones and other proteins.
39 citations
TL;DR: The results may suggest the correlation between binding of D-amino acid to peptidoglycan and inhibition of formation of the bound form of lipoprotein.
Abstract: Certain D-amino acids, such as D-methionine and D-cystine, were incorporated into cells of Escherichia coli under conditions inhibiting protein and cell wall synthesis. Part of the radioactivity of D-14C-amino acids incorporated into the cells was found in the isolated cell wall peptidoglycan. A covalent linkage between the amino group of the D-amino acids and the peptidoglycan was presumed to be the main cause of the binding of the D-amino acids to peptidoglycan, because the amino group of the D-amino acids in the incorporation product was substituted. Whether the carboxyl terminus was substituted was unknown. The formation of the D-amino acid-peptidoglycan linkage was insensitive to beta-lactam antibiotics such as benzylpenicillin and ampicillin (500 micrograms/ml) and therefore was not due to the reaction of DD-transpeptidation which is involved in the biosynthesis of peptidoglycan. The D-amino acids also strongly inhibited the formation of peptidoglycan-bound lipoprotein in the E. coli cells. The results may suggest the correlation between binding of D-amino acid to peptidoglycan and inhibition of formation of the bound form of lipoprotein.
32 citations
TL;DR: The finding of these glycopeptides in urine shows that mucolipidosis I is characterized by a general "glycoprotein-specific" sialidase deficiency, and the possibility of the existence of a human endo-alpha-N-acetylgalactosaminidase is discussed.
Abstract: Sialyl-glycopeptides containing an O-glycosidically linked tetrasaccharide chain were obtained from the urine of a patient suffering from mucolipidosis I. Isolation of these compounds was achieved by gel filtration, ion-exchange chromatography and preparative paper chromatography. Their structures were determined by a combination of carbohydrate and amino acid analysis, dansylation, periodate oxidation, methylation studies, enzymatic hydrolysis and 1H-NMR spectroscopy, to be as follows:
wherein R = peptide linked through -Thr-, -Ser-, -Ser-Thr- or -Thr-Ser-.
The finding of these glycopeptides in urine shows that mucolipidosis I is characterized by a general “glycoprotein-specific” sialidase deficiency. The possibility of the existence of a human endo-α-N-acetylgalactosaminidase is discussed.
25 citations
TL;DR: Procedures for the stepwise addition of one or more deoxyribonucleotide residues to the 3' end of an oligodeoxy ribonucleoside phosphate acceptor using commercially available terminal deoxynucleotidyl transferase is described.
Abstract: Procedures for the stepwise addition of one or more deoxyribonucleotide residues to the 3' end of an oligodeoxyribonucleoside phosphate acceptor using commercially available terminal deoxynucleotidyl transferase is described. 2-80 nmol of acceptors with a chain length of four, five or nine monomer units were elongated with a single 2'-deoxyribonucleoside 5'-triphosphate in yields of 20-30%. The monomers carried no protecting groups and were used both radioactively labelled and unlabelled. The elongated oligodeoxynucleoside phosphates were isolated by reverse-phase (Nucleosil C18) high-performance liquid chromatography or paper chromatography. The isolated products were sequenced by the fingerprint method. Advantages and disadvantages of this new methodology for the enzymatic synthesis of defined oligodeoxynucleotides are discussed.
18 citations
TL;DR: Three phosphate-containing sialyloligosaccharides isolated from normal human urine may indicate the existence of a novel, as yet unrevealed pathway in the metabolism of complex carbohydrates.
Abstract: Three phosphate-containing sialyloligosaccharides were isolated from normal human urine using charcoal adsorption, gel-filtration chromatography, ion-exchange chromatography and paper chromatography Studies including gas-liquid chromatography of monosaccharide and disaccharide derivatives, methylation analysis, phosphate determination, ion-exchange chromatography and glycosidase and phosphatase treatments indicated the following three structures for the compounds isolated: NeuAc(α2–6)Gal(β1–4)GlcNAc(α)-P; NeuAc(α2–3)Gal(β1–4)GlcNAc(α)-P; NeuAc(α2–3)Gal(β1–3)GalNAc(α)-P
These sialyloligosaccharide 1-phosphates represent a novel class of oligosaccharides Their oligosaccharide chains are identical with the common sialyloligosaccharide end groups of glycoproteins and glycolipids The excretion of these compounds in normal human urine may indicate the existence of a novel, as yet unrevealed pathway in the metabolism of complex carbohydrates
16 citations
TL;DR: It is concluded that transfer of Glc3Man9GlcNAc2 to protein is followed by excision of glucose and probably one or two mannose residues, followed by further mannosylation and in some cases also methylation of oligosaccharides, which represents a novel reaction in the processing of N-linked oligosACcharides.
TL;DR: In this paper, the effect of the vapor phase and other special influences on thin-layer chromatography have been investigated and the relationship of time vs. developing distance and flow rate vs. efficiency using a planar arrangement of the thin layer was made.
Abstract: The effect of the vapor phase and other special influences on thin-layer chromatography have been investigated. Comparisons were made of the relationships of time vs. developing distance and flow rate vs. efficiency using a planar arrangement of the thin-layer. Covering the layer facilitates the reproducibility and of the migration front but the most effective optimization step for thin-layer chromatography is provided by forced-flow of the mobile phase. It is suggested that planar chromatography with a covered sorbent layer and using a pressurised solvent stream should be calledforced-flow thin-layer chromtatography.
TL;DR: The results suggest that the lipid-bound saccharides are involved in the biosynthesis of the extracellular polysaccharides.
Abstract: Summary: The incubation of an enzyme preparation of Agrobacterium tumefaciens with UDP-[14C]Glc led to the formation of radioactive substances soluble in chloroform/methanol/water (1 : 1 : 0·3, by vol.). These substances had properties similar to the polyprenol diphosphate saccharides. The lipid moiety appears to be an unsaturated polyprenol. Mild acid hydrolysis of the substances liberated five water-soluble products which were separated by TLC and characterized by borohydride reduction-acid hydrolysis, partial acid hydrolysis, methylation analysis, TLC and paper chromatography and paper electrophoresis. The products behaved like: galactose; the disaccharide Glc(β1-3)Gal; the tetrasaccharide Glc(β1-4)Glc(β1-4)Glc(β1-3)Gal; an octasaccharide and a pyruvylated octasaccharide. The last two substances were compared by paper electrophoresis, TLC and methylation analysis with an octasaccharide obtained by the action of a hydrolytic enzyme on the exopolysaccharide of Agrobacterium tumefaciens. The octasaccharide and the pyruvylated octasaccharide from the lipid behaved like those obtained by enzyme action on the A. tumefaciens exopolysaccharide. These results suggest that the lipid-bound saccharides are involved in the biosynthesis of the extracellular polysaccharides.
Journal Article•
TL;DR: The data suggest that depressed inflammatory responses at sites of neoplasms may result in part from release of small, potent inhibitors of leukotaxis from tumors themselves.
Abstract: Lysate from T-241 murine fibrosarcoma cells contains a low-molecular-weight ( M r < 1000), heat-stable peptide factor which has antichemotactic activity for both macrophages and polymorphonuclear leukocytes in vitro . The tumor factor was partially purified from an alcohol extract of the fibrosarcomas by gel filtration, anion exchange chromatography, and paper chromatography successively. This factor inhibits both the hydrolytic cleavage of the peptide attractant N -formylmethionylleucylphenylalanine by polymorphonuclear leukocytes and the methylation of both protein carboxyl groups and membrane phospholipids. Furthermore, the factor does not appear to compete with N -formylmethionylleucylphenylalanine for its receptor. The tumorderived material, therefore, affects biochemical reactions believed to have roles in the expression of an adequate leukotactic response. These data suggest that depressed inflammatory responses at sites of neoplasms may result in part from release of small, potent inhibitors of leukotaxis from tumors themselves.
TL;DR: Oligosaccharides, which are O- and N-glycosidically linked on salivary glycoprotein from the edible bird's nest of chinese swallows, were released by alkaline borohydride treatment of the asialoglycoproteins and fractionated by gel chromatography.
Abstract: Oligosaccharides, which are O- and N-glycosidically linked on salivary glycoproteins from the edible bird's nest of chinese swallows, were released by alkaline borohydride treatment of the asialoglycoproteins and fractionated by gel chromatography. Fract. VN1 (oligosaccharides greater than 2 000 dalton) apparently represented a mixture of saccharides derived from complex, N-glycosidically linked glycans (molar ratio Man/GlcNAc/Gal 3:4:8), while fractions VN2 (tetra- to hexasaccharides), VN3 (trisaccharide) and VN4 (disaccharide) were free of mannose, but did contain all the N-acetylgalactosamine released from the protein as its alditol. Oligosaccharides in Fract. VN2 and VN4 were purified by high-performance liquid chromatography, paper chromatography and thin-layer chromatography, methylated and analysed after total or partial acid hydrolysis by gas-liquid chromatography-mass spectrometry. The structures of a hexasaccharide in Fract. VN2/6 and of a tetrasaccharide in fraction VN2/4 were finally established after methylation through direct-probe mass spectrometry: Gal(1----4)GlcNAc(1----3)Gal(1----4)GlcNAc(1----3)Gal(1----3)GalNAc- ol and Gal(1----4)GlcNAc(1----6)[Gal(1----3)]GalNAc-ol. Mass spectrometrical and gas-chromatographical data obtained for a disaccharide in Fract. VN4 were identical with those for Gal(beta 1----3)GalNAc-ol.
01 Jan 1984
TL;DR: The amino acid sequences of 39 tryptic peptides from carboxymethylated mitochondrial aspartate amino- transferase from pig heart muscle were analyzed and accounted for 318 of the total 401 amino acid residues in the protein subunit.
Abstract: The amino acid sequences of 39 tryptic peptides from carboxymethylated mitochondrial aspartate amino- transferase from pig heart muscle were analyzed. The peptides were purified by gel filtration, ion exchange column chromatography, paper chromatography, and high voltage paper electrophoresis, and their sequences were examined by manual Edman degradation, carbox- ypeptidase digestion, and fragmentation with thermol- ysin or chymotrypsin. These peptides accounted for 318 of the total 401 amino acid residues in the protein subunit.
TL;DR: 99mTc-dimercaptosuccinic acid /DMSA/ is one of the most favourable agents used for renal scintigraphy and was prepared in two different ways: by direct labelling of tin/II/-dimercaptic acid and by ligand exchange reaction at tracer concentration of technetium under acidic condition.
Abstract: 99mTc-dimercaptosuccinic acid /DMSA/ is one of the most favourable agents used for renal scintigraphy. This radiopharmaceutical was prepared in two different ways: by direct labelling of tin/II/-dimercaptosuccinic acid and by ligand exchange reaction from99mTc-gluconate at tracer concentration of technetium under acidic condition. The complexes formed were compared using paper chromatography, thin layer chromatography, gel filtration and electrophoresis. Their biodistribution was studied on rats.
TL;DR: Preparative paper chromatography is proposed as a suitable method for purification of Xylenol Orange (XO) and the co-existence of ML and M(2) L complexes proved by use of the program DALSFEK.
TL;DR: Calf kidney has been used as a tissue source for the isolation of cobalamin analogues, which are defined here as cobalt-containing compounds of distinctive chromatographic behavior that are extractable from tissues by methods conventionally used to extract cobalamins and which are more active with R-protein as binder than intrinsic factor and are relatively less active in microbiologic assays.
TL;DR: A double-label, double-substrate incubation technique has been developed and used to study the conversion of progesterone to testosterone in testes extracts from incompletely virilized males and represents an improvement in time, reproducibility, and simplicity when compared to techniques based on thin layer or paper chromatography.
Abstract: A double-label, double-substrate incubation technique has been developed and used to study the conversion of progesterone to testosterone in testes extracts from incompletely virilized males. The procedure involves separation of the microsomes from a testicular homogenate, incubating the microsomes with 1 microM [7-3H] progesterone, 1 microM 17-hydroxy[4-14C]progesterone, and 0.25 mM NADPH in pH 7.4 phosphate buffer at 37 degrees C. Steroid precursors and products are separated by column chromatography on Sephadex LH-20 with a solvent system of isoctane:ethyl acetate:methanol (4:1:1 by volume). These procedures can be completed in 2 days, and thus the method represents an improvement in time, reproducibility, and simplicity when compared to techniques based on thin layer or paper chromatography. The method has been used to distinguish the biochemical abnormality in three cases with XY sex chromatin, posterior labial fusion, clitoromegaly, and hypospadias. The abnormalities identified were: Case 1, no defect in testosterone synthesis (probable androgen insensitivity); Case 2, 17-ketosteroid reductase deficiency; and Case 3, steroid-17, 20-lyase deficiency.
TL;DR: In this article, the authors discuss thin layer chromatography and paper chromatography, which together are composed of "flat-bed" or "planar" chromatography; they provide qualitative analytical information, and with optimization of techniques and materials, can give quantitative data as well.
Abstract: Publisher Summary This chapter discusses thin layer chromatography (TLC) and paper chromatography (PC), which together is composed of “flat-bed” or “planar” chromatography, are the easiest of all chromatographic methods to perform, and they require only simple and inexpensive equipment. Many of the techniques and principles of TLC and PC are the same. They both provide qualitative analytical information, and, with optimization of techniques and materials, can give quantitative data as well. An initial zone of mixture is placed near one end of the stationary phase, a thin layer or paper sheet; the sample is dried; and the end of the stationary phase with the initial zone is placed into a mobile phase, usually a mixture of pure solvents, inside a closed chamber. The components of the mixture migrate at different rates during the movement of the mobile phase through the stationary phase, which is termed the development of the chromatogram. When the mobile phase has moved an appropriate distance, the stationary phase is removed, the mobile phase is rapidly dried, and the zones are detected by application of a suitable visualization reagent. Sample collection, preservation and purification are common problems in both TLC and PC. Detection is most simple when the compounds of interest are naturally colored or fluorescent or absorb ultraviolet (UV) light.
TL;DR: Purified mitochondria from rat liver were found to contain protein synthesis inhibitors, that could be extracted by disruption of mitochondrial membranes and fractionated by gel filtration into two fractions of low and high molecular weight.
Abstract: Purified mitochondria from rat liver were found to contain protein synthesis inhibitors, that could be extracted by disruption of mitochondrial membranes and fractionated by gel filtration into two fractions of low and high molecular weight. Small size inhibitors were also released from the latter peak by high ionic strength followed by gel filtration. Both types of factors inhibit incorporation of radioactive amino acids into protein by liver cytoplasmic polysomes programmed with endogenous mRNA or poly U, and by rabbit reticulocyte lysates programmed with added globin mRNA and by incubations of Walker carcinoma cells. They decrease to the same level the cytoplasmic synthesis of proteins for the mitochondrial and extra-mitochondrial compartments in intact cells, but do not appear to inhibit substantially endogenous mitochondrial protein synthesis. Inhibitors were purified by paper chromatography and reverse phase high performance liquid chromatography into fractions which block with the same kinetics the incorporation of [14]leucine and [35]methionine into protein in systems able to initiate protein synthesis, such as reticulocyte lysates or intact cells, but differ in this respect in incubations of liver ribosomes where re-binding of mRNA is a limiting step. Some of these factors behave as oligopeptides that are assumed to inhibit in vitro primarily the initiation stage but whose function in vivo is still undetermined.
TL;DR: The presence of an enzyme involved in ATP synthesis by transfer of phosphate from thiamine triphosphate to ADP in maize germ axis was indicated by the assays on partially purified (27-fold) enzyme with luciferase method, spectrophotometric assay with hexokinase and glucose-6-phosphates dehydrogenase, and paper chromatography.
Abstract: The presence of an enzyme involved in ATP synthesis by transfer of phosphate from thiamine triphosphate to ADP in maize germ axis was indicated by the assays on partially purified (27-fold) enzyme with luciferase method, spectrophotometric assay with hexokinase and glucose-6-phosphate dehydrogenase, and paper chromatography. Optimal activity was found at pH 9.0. The enzyme was heat-labile SH-enzyme, and its activity required the presence of Mg2+.
TL;DR: Procedures for the identification of decoction of Alismatis Rhizoma (rhizome of Alisma orientale JUZEPCZUK or related species) by cellulose acetate membrane electrophoresis, paper partition chromatography, and thin-layer chromatography were developed.
Abstract: Procedures for the identification of decoction of Alismatis Rhizoma (rhizome of Alisma orientale JUZEPCZUK or related species) by cellulose acetate membrane electrophoresis, paper partition chromatography, and thin-layer chromatography were developed. These methods are due to the presence of a characteristic new component.
TL;DR: In addition to characterizing the immune chromatography procedure, the possible applications of chromatography assays for the quantitation of other types of molecular binding interactions are discussed.