scholarly journals The structure of carbohydrate unit B of porcine thyroglobulin

1981 ◽  
Vol 195 (3) ◽  
pp. 701-713 ◽  
Author(s):  
K Yamamoto ◽  
T Tsuji ◽  
T Irimura ◽  
T Osawa
Keyword(s):  
Sialic Acid ◽  
Concanavalin A ◽  
Deae Cellulose ◽  
Structural Features ◽  
The Other ◽  
Methylation Analysis ◽  
Complex Type ◽  
Mannose Residue ◽  
Sugar Chains ◽  
Carbohydrate Unit

The oligosaccharide fraction was obtained from porcine thyroglobulin by hydrazinolysis. Four fractions of unit B-type oligosaccharides were purified by successive chromatographies on columns of DEAE-cellulose and concanavalin A-Sepharose, and their structures were investigated by the combination of endo- and exo-glycosidase digestions, methylation analysis and Smith degradation. From the results of these studies, the structures of the unit B oligosaccharides were proposed to be as follows: see formula in text. Thus the glycoprotein was found to have triantennary and biantennary complex-type oligosaccharides as acidic sugar chains. Concerning the triantennary oligosaccharides, the following structural features were shown: (1) the sialic acid residues were not localized on certain specific branches but distributed on all three branches; (2) however, alpha (2 leads to 3)-linked sialic acid residues were exclusively located on the terminal of the branch arising from C-4 of the branching alpha-mannose residue, whereas alpha (2 leads to 6)-linked sialic acid residues occupied terminals of the other branches; (3) the outer branching alpha-mannose residue was attached to C-3 or C-6 of an inner branching beta-linked mannose residue, and both types were observed to exist.

scholarly journals Structure of carbohydrate unit A or porcine thyroglobulin

1981 ◽  
Vol 195 (3) ◽  
pp. 691-699 ◽  
Author(s):  
T Tsuji ◽  
K Yamamoto ◽  
T Irimura ◽  
T Osawa
Keyword(s):  
Column Chromatography ◽  
Core Structure ◽  
Methylation Analysis ◽  
Complex Type ◽  
Form Complex ◽  
Sugar Chains ◽  
Carbohydrate Unit ◽  
Terminal Mannose

The unit A-type glycopeptides were purified from porcine thyroglobulin by Pronase digestion followed by chromatography on a DEAE-Sephadex A-25 column. These glycopeptides were separated into five fractions (UA-I, -II, -IV and -V) by Dowex 50W (X2) column chromatography. Fractions UA-I, -II, -III, -IV and -V were found to have the compositions (Man)9(GlcNAc)2-Asn, (Man)8(GlcNAc)2-Asn, (Man)7(GlcNAc)2-Asn, (Man)6(GlcNAc)2-Asn and (Man)5(GlcNAc)2-Asn respectively. The structures of these five fractions were investigated by the combination of exo- and endo-glycosidase digestions, methylation analysis. Smith periodate degradation and acetolysis. The results showed that fraction UA-V had the simplest structure: see formula in text. The larger glycopeptides (fractions UA-I, -II, -III and -IV) contained additional mannose residues alpha (1 leads to 2)-linked to the terminal mannose residues in the above core structure. These unit A-type glycopeptides appear to be biosynthetic intermediates that are to be processed to form complex-type glycopeptides (unit B-type sugar chains).

Studies on the к-casein complex: II. The isolation of a sialic acid-containing fraction by disrupting the complex at low pH in the presence of sodium chloride

1965 ◽  
Vol 32 (1) ◽  
pp. 57-63 ◽  
Author(s):  
R. Beeby
Keyword(s):  
Sialic Acid ◽  
Sodium Chloride ◽  
Deae Cellulose ◽  
Low Ph ◽  
The Other ◽  
Trichloracetic Acid ◽  
Complex Ii ◽  
Total Sialic Acid

SummaryWhen crude к-casein was precipitated at pH 3 in the presence of 0·4m-NaCl the supernatant contained up to 80% of the total sialic acid but no detectable cystine or cysteine. Two fractions were obtained from this supernatant by chromatography on DEAE cellulose; one containing 4–6% sialic acid and the other only onetenth of this amount.Most of the sialic acid of the sialic acid-rich fraction was soluble in 12% trichloracetic acid following treatment with rennin. It is suggested that the glycopeptide released by the action of the enzyme on casein originates from this fraction.

scholarly journals Carbohydrate structure of human pancreatic elastase 1

1991 ◽  
Vol 278 (2) ◽  
pp. 505-514 ◽  
Author(s):  
P Wendorf ◽  
D Linder ◽  
A Sziegoleit ◽  
R Geyer
Keyword(s):  
Gel Filtration ◽  
Methylation Analysis ◽  
Complex Type ◽  
Pancreatic Elastase ◽  
Blood Group A ◽  
Elastase 1 ◽  
Glycosylation Sites ◽  
Group A ◽  
Sugar Chains ◽  
Pancreatic Elastase 1

Human pancreatic elastase 1 (E1) is a glycoprotein containing two potential N-glycosylation sites, one of which carries a carbohydrate moiety [Wendorf, Geyer, Sziegoleit & Linder (1989) FEBS Lett. 249, 275-278]. In order to study its glycosylation, glycoprotein isolated from post-mortem pancreas tissue of 75 donors was digested with trypsin. Oligosaccharides were liberated from resulting glycopeptides by treatment with peptide-N4-(N-acetyl-beta-glycosaminyl)-asparagine amidase F, radiolabelled by reduction with KB3H4 and separated by h.p.l.c. and gel filtration. Major oligosaccharide alditol fractions, representing 67.8 mol% of total glycans, were characterized by methylation analysis and sequential degradation with exoglycosidases. The results revealed that about two-fifths of the partially truncated, mainly biantennary, complex-type glycans found comprised blood group A, B, Lea (or X), difucosyl A or difucosyl B determinants, which could be assigned to lactosamine antennae linked to Man(alpha 1-3)- residues of the sugar chains.

scholarly journals Glycophorin A interferes in the agglutination of human erythrocytes by concanavalin A Explanation of the requirement for enzymic predigestion

1987 ◽  
Vol 241 (2) ◽  
pp. 505-511 ◽  
Author(s):  
S M Gokhale ◽  
N G Mehta
Keyword(s):  
Sialic Acid ◽  
Correlation Coefficient ◽  
Concanavalin A ◽  
Binding Sites ◽  
Band 3 ◽  
Human Erythrocytes ◽  
The Other ◽  
Glycophorin A ◽  
Con A

Human erythrocytes become agglutinable with concanavalin A (Con A) after treatment with various proteinases or neuraminidase. The extent of agglutinability achieved with different enzymes is, however, different: Pronase, papain, trypsin, neuraminidase and chymotrypsin enhance the agglutinability in decreasing order, the last being barely effective. The actions of the enzymes on band 3, the Con A receptor, do not correlate with their abilities to increase the agglutinability: Pronase, papain and chymotrypsin cleave the protein, but not trypsin or neuraminidase. No significant differences are found in the number of Con A-binding sites or the affinities for the lectin between the normal and trypsin- or Pronase-treated cells. Thus the receptor does not seem to play a role in determining the Con A-agglutinability of erythrocytes. On the other hand, the cleavage of glycophorins, especially glycophorin A, and the release of sialic acid (in the peptide-bound form) are well-correlated with the enhancement in agglutination after the action of proteinases. The release of sialic acid by graded neuraminidase digestion and the increase in Con A-agglutinability show a correlation coefficient of 0.88. The major inhibitory role of glycophorin A in the process is indicated by the agglutination of En(a) heterozygous erythrocytes; the cells, known to bear about 50% glycophorin A molecules in their membrane, are agglutinated approximately half as well without proteolysis as are the trypsin-treated cells. Possible mechanisms by which glycophorin A could affect Con A-mediated agglutination are discussed.

scholarly journals Postnatal changes in N-linked oligosaccharides of glycoproteins in rat liver

1988 ◽  
Vol 253 (1) ◽  
pp. 59-66 ◽  
Author(s):  
S Kato ◽  
S Oda-Tamai ◽  
N Akamatsu
Keyword(s):  
Rat Liver ◽  
Concanavalin A ◽  
Post Partum ◽  
Deae Cellulose ◽  
Complex Type ◽  
Processing Enzymes ◽  
Significant Difference ◽  
Neonatal Liver ◽  
Old Rats ◽  
Rat Livers

[3H]Mannose-labelled glycopeptides in the slices of livers from neonatal and 1-, 2-, 3- and 5-week-old rats were characterized by column chromatographies on Sephadex G-50 and concanavalin A-Sepharose and by endo-beta-N-acetylglucosaminidase H digestion. The proportion of complex-type glycopeptides was increased with time until 2 weeks post partum and then returned to the neonatal level. This was mainly due to the increased proportion of concanavalin A-bound (biantennary) species. These changes were accompanied by consistent changes in the activities of processing enzymes in liver microsomal fraction, especially of N-acetylglucosaminyltransferase I. Complex-type glycopeptides from neonatal and 2- and 5-week-old rat livers were further characterized by column chromatographies on Bio-Gel P-6 and DE 52 DEAE-cellulose in combination with neuraminidase digestion. No significant difference was found between concanavalin A-bound species from neonatal liver and those from liver 5 weeks post partum, most of which were sialylated. Concanavalin A-bound species 2 weeks post partum were comparatively smaller in size and less sialylated. On the other hand, there was no significant difference among concanavalin A-unbound species from the three different sources, most of which were sialylated. Since glycoproteins from regenerating rat liver also contain a higher proportion of complex-type oligosaccharides, as previously reported, such changes in N-linked oligosaccharides of glycoproteins may be related to control of the growth of liver cells.

scholarly journals The multiple forms of α-D-mannosidase in human plasma

1979 ◽  
Vol 179 (3) ◽  
pp. 583-592 ◽  
Author(s):  
S Hirani ◽  
B Winchester
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
The Other ◽  
Multiple Forms ◽  
Simple Correlation ◽  
Multiple Components

The acidic alpha-D-mannosidase in human plasma closely resembles liver acidic alpha-D-mannosidase in its affinity for concanavalin A-Sepharose, molecular weight and resolution into multiple components on DEAE-cellulose. A combination of chromatography on concanavalin A-Sepharose and gel filtration on Sephadex G-200 and Sepharose 6B suggests that four forms of intermediate alpha-D-mannosidase, which differ either in their molecular weight of affinity for concanavalin A, exist in human plasma. A practical classification and nomenclature for the multiple forms of intermediate alpha-D-mannosidase in plasma based on molecular weight and affinity for concanavalin A is proposed. Multiple forms of intermediate alpha-D-mannosidase were also observed by ion-exchange chromatography on DEAE-cellulose, but there was not a simple correlation between these forms and those obtained with the other separation procedures. The form of intermediate alpha-D-mannosidase least abundant in plasma, approx. 7% of the activity, has very similar properties to the neutral alpha-D-mannosidase in human liver. In contrast, the other three forms of intermediate alpha-D-mannosidase, which account for over 90% of the activity, do not appear to be present in liver, except perhaps in trace amounts.

scholarly journals A unique heparan sulfate in the nuclei of hepatocytes: structural changes with the growth state of the cells.

1986 ◽  
Vol 102 (2) ◽  
pp. 587-599 ◽  
Author(s):  
N S Fedarko ◽  
H E Conrad
Keyword(s):  
Heparan Sulfate ◽  
Nuclear Membrane ◽  
Structural Changes ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Structural Features ◽  
The Other ◽  
Growth State ◽  
Before And After ◽  
Heparan Sulfates

Growing and confluent cultures of a rat hepatocyte cell line were labeled with 35SO4(2-) and the heparan sulfate in the culture medium, the pericellular matrix, the nucleus, the nuclear outer membrane, and the remaining cytoplasmic pool was purified by DEAE-cellulose chromatography. The heparan sulfate in all pools from the confluent cells was bound more strongly on the DEAE-cellulose column than the corresponding pools from the growing cells. Gel filtration of each pool before and after beta-elimination showed that the heparan sulfate from the nuclear and nuclear membrane pools was composed of primarily free chains, whereas the heparan sulfate in all of the other pools was a mixture of proteoglycans and free chains. The heparan sulfate in each pool was cleaved with nitrous acid to obtain mixtures of di- and tetrasaccharides. Analysis of these mixtures showed that the structural features of the heparan sulfates in each pool were different and were altered significantly when the growing cells became confluent. The nuclear-plus-nuclear membrane pools represented 6.5% and 5.4% of the total cell-associated heparan sulfate in the growing cells and the confluent cells, respectively. The structural features of the heparan sulfate in the two nuclear pools were very similar to each other, but were markedly different from those of the heparan sulfate from the other pools or from any previously described heparan sulfate or heparin. The most unusual aspect of these structures was the high content of beta-D-glucuronosyl(2-SO4)----D-glucosamine-N,O-(SO4)2 disaccharide units in these sequences. The mode of biosynthesis and delivery of these unusual sequences to the nucleus and the potential significance of these observations are discussed.

scholarly journals Type analysis of the oligosaccharide chains on microheterogeneous components of bovine pancreatic DNAase by the lectin-nitrocellulose sheet method

1989 ◽  
Vol 257 (1) ◽  
pp. 43-49 ◽  
Author(s):  
S Kijimoto-Ochiai ◽  
Y U Katagiri ◽  
T Hatae ◽  
H Okuyama
Keyword(s):  
Sialic Acid ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Complex Type ◽  
Type Analysis ◽  
Mannose Residue ◽  
Hybrid Type ◽  
Oligosaccharide Chain ◽  
Ricinus Communis Agglutinin ◽  
High Mannose

The oligosaccharide chains of microheterogeneous bovine pancreatic DNAases were characterized by the lectin-nitrocellulose sheet method. The active fractions of the DNAases from column chromatography showed four major and several minor spots on a two-dimensional polyacrylamide gel. They were transferred on to nitrocellulose sheets and treated with glycosidases (neuraminidase, endo-beta-N-acetyl glucosaminidase H or F, or peptide N-glycosidase F) and treated with peroxidase-coupled lectins (concanavalin A, Ricinus communis agglutinin or wheat-germ agglutinin). From the results, the most probable oligosaccharide types were proposed to be as follows: the four major spots contained components which had high-mannose type or hybrid-type oligosaccharides, such as those susceptible to endo-beta-N-acetylglucosaminidase H. In addition, spot 1 contained a complex-type biantennary oligosaccharide without sialic acid and spot 3 contained a tri- or tetra-antennary complex-type oligosaccharide with sialic acid. The component corresponding to spot 2 had a hybrid-type oligosaccharide chain with a ‘bisecting’ acetylglucosamine, linked 1-4 to the beta-mannose residue of the trimannosyl core, and the component corresponding to spot 4 had a high-mannose-type oligosaccharide chain.

scholarly journals The wall teichoic acids of Lactobacillus plantarum N.I.R.D. C106. Location of the phosphodiester groups and separation of the chains

1971 ◽  
Vol 124 (3) ◽  
pp. 449-460 ◽  
Author(s):  
A. R. Archibald ◽  
Hilary E. Coapes
Keyword(s):  
Lactobacillus Plantarum ◽  
Concanavalin A ◽  
Teichoic Acid ◽  
The Other ◽  
Hydroxyl Group ◽  
Methylation Analysis ◽  
Teichoic Acids ◽  
Wall Teichoic Acids ◽  
Adjacent Unit

1. The identities of the component glycerol glucosides of the wall teichoic acids of Lactobacillus plantarum N.I.R.D. C106 have been confirmed by methylation analysis. These glucosides are α-d-glucopyranosyl-(1→1)-l-glycerol, α-d-glucopyranosyl-(1→2)-α-d-glucopyranosyl-(1→1)-l-glycerol and α-d-glucopyranosyl-(1→3)-α-d-glucopyranosyl-(1→1)-l-glycerol. 2. These units are connected by phosphodiester groups attached to the 3(l)-hydroxyl group of glycerol and the 6-hydroxyl group of the non-reducing terminal glucose residues in the adjacent unit. 3. Concanavalin A forms a precipitate with the teichoic acid and the material so precipitated contains only the α-d-glucopyranosyl-(1→2)-α-d-glucopyranosyl -(1→1)-l-glycerol component. This unit is therefore present in a homogeneous polymer so that the teichoic acid is a mixture of this and of other possibly homogeneous chains containing the other two components.

Quo Vadis Tyrannosaurus?: The Future of Dinosaur Studies

1989 ◽  
Vol 2 ◽  
pp. 175-183 ◽  
Author(s):  
Daniel J. Chure
Keyword(s):  
Public Relations ◽  
Earth Science ◽  
Structural Features ◽  
The Other ◽  
Canada Geese ◽  
New Genera ◽  
Research Activity ◽  
Detailed Understanding ◽  
The Public ◽  
Quo Vadis

“Although I work a lot with fossils in my own research on fishes, I do not care to be called a paleontologist; and I am turned off by many aspects of the public-relations hoopla surrounding paleontology, especially dinosaurs…. One could easily argue that the schools' fascination with dinosaurs might also detract from the other aspects of earth science and biological science and, in the end, weaken paleontology's image as an activity for hard-nosed grown-ups.”K.S. Thomson, 1985: p. 73“Let dinosaurs be dinosaurs. Let the Dinosauria stand proudly alone, a Class by itself. They merit it. And let us squarely face the dinosaurness of birds and the birdness of the Dinosauria. When the Canada geese honk their way northward, we can say: “The dinosaurs are migrating, it must be spring!”R.T. Bakker, 1986: p. 462It is a now oft-repeated statement that we are in the Second Golden Age of dinosaur studies. This may at first seem to be yet another overstatement by dinosaur fanatics; in fact, it is substantiated on a number of fronts. Research activity is certainly at an all-time high, with resident dinosaur researchers on every continent (except Antarctica) and dinosaurs known from every continent (including Antarctica). This activity has resulted in a spate of discoveries, including not only new genera and species, but entirely new types of dinosaurs, such as the segnosaurs. Well-known groups are producing surprises, such as armored sauropods and sauropods bearing tail clubs. Good specimens of previously named genera are revealing unsuspected structural features that almost defy explanation, as in the skull of Oviraptor. However, dinosaur studies extend far beyond the traditional emphasis on dinosaur morphology, and encompass paleobiogeography, paleoecology, taphonomy, physiology, tracks, eggs, histology, and extinction, among others. In some cases, several of these studies can be applied to a single taxon or locality to give us a fairly detailed understanding of the paleobiology of some species.

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