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Query: UMLS:C0027960 (mole)
 21,279 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Human immunoglobulin G (IgG) is unique among serum glycoproteins because it contains more than 30 different biantennary complex-type asparagine-linked oligosaccharides. This extremely high microheterogeneity is probably produced because human individuals have a series of B cell clones equipped with different sets of glycosyltransferases. Despite this complex composition, IgG samples purified from whole human sera have the same mole ratios of oligosaccharides, indicating that the ratio of B cell clones synthesizing IgGs with different sugar chains is constant in healthy individuals. We found that the glycosylation patterns of whole serum IgGs obtained from patients with rheumatoid arthritis (RA) are quite different from those of whole serum from healthy individuals. Structural studies of the oligosaccharides revealed that the sugar chains of the IgGs obtained from patients with RA are depleted of the beta-galactose residues. The sugar chains of transferrin from patients with RA are fully galactosylated. Therefore the galactose deletion from IgG is probably brought about by a decrease in galactosyltransferase activity in B cells rather than by degradation by galactosidase during circulation. Enzymic study revealed that human B cells contain various beta-galactosyltransferases which form the Gal(beta 1-4)GlcNAc groups in the sugar chains of different glycoproteins. Among these enzymes, abnormality in patients with RA was found only in the one that transfers beta-galactose residues specifically to degalactosylated IgG. This enzyme showed lower affinity toward UDP-Gal in B cells of patients with RA than that in healthy individuals.
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PMID:Function and pathology of the sugar chains of human immunoglobulin G. 279 53

Solvent and solvent proton dependent steps involved in the mechanism of the enzyme galactose oxidase have been examined. The deuterium kinetic solvent isotope effect (KSIE) on the velocity of the galactose oxidase catalyzed oxidation of methyl beta-galactopyranoside by O2 was measured. Examination of the thermodynamic activation parameters for the reaction indicated that the isotope effect was attributable to a slightly less favorable delta H value, consistent with a KSIE on proton transfer. A detailed kinetic analysis was performed, examining the effect of D2O on the rate of reaction over the pH range 4.8-8.0. Both pL-rate profiles exhibited bell-shaped curves. Substitution of D2O as solvent shifted the pKes values for the enzymic central complex: pKes1 from 6.30 to 6.80 and pKes2 from 7.16 to 7.35. Analysis of the observed shifts in dissociation constants was performed with regard to potential hydrogenic sites. pKes1 can be attributed to a histidine imidazole, while pKes2 is tentatively assigned to a Cu2+-bound water molecule. A proton inventory was performed (KSIE = +1.55); the plot of kcat vs. mole fraction D2O was linear, indicating the existence of a single solvent-derived proton involved in a galactose oxidase rate-determining step (or steps). The pH dependence of CN- inhibition was also examined. The Ki-pH profile indicated that a group ionization, with pKa = 7.17, modulated CN- inhibition; Ki was at a minimum when this group was in the protonated state. The inhibition profile followed the alkaline limit of the pH-rate profile for the enzymic reaction, suggesting that the group displaced by CN- was also deprotonating above pH 7.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Solvent and solvent proton dependent steps in the galactose oxidase reaction. 282 Apr 69

Structures of the sugar moieties of gamma-glutamyl transpeptidases purified from the kidney and the liver of rat, mouse, and cattle were studied after being chemically released as oligosaccharides. The results indicated that both organ-specific and species-specific differences exist in the sugar chains of the enzyme. Comparative studies of sugar chains of the heavy and light subunits of the rat kidney enzyme revealed that high mannose-type sugar chains are found only in the heavy subunit. By the same analysis of the oligosaccharide fractions obtained from four isozymic forms of the rat kidney enzyme, it was found that all these enzymes contain 2 mole of neutral sugar chains but different numbers of acidic sugar chains in one molecule. Comparative studies of oligosaccharides obtained from the enzymes purified from rat AH-66 hepatoma and from normal rat liver revealed that more than 40% of the sugar chains of the hepatoma enzyme contain bisecting N-acetylglucosamine residues which are not found in those of the liver enzyme. By making use of the structural changes associated with malignant transformation, a new diagnostic method or hepatoma was developed. In principle, the method consists of affinity chromatography of the desialylated serum enzyme using an erythroagglutinating lectin agarose column.
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PMID:[Structural changes in the sugar chains of gamma-glutamyltranspeptidase by malignant transformation]. 287 94

When the heat-activated chloroplast F1 ATPase hydrolyzes [3H, gamma-32P]ATP, followed by the removal of medium ATP, ADP, and Pi, the enzyme has labeled ATP, ADP, and Pi bound to it in about equal amounts. The total of the bound [3H]ADP and [3H]ATP approaches 1 mol/mol of enzyme. Over a 30-min period, most of the bound [32P]Pi falls off, and the bound [3H]ATP is converted to bound [3H]ADP. Enzyme with such remaining tightly bound ADP will form bound ATP from relatively high concentrations of medium Pi with either Mg2+ or Ca2+ present. The tightly bound ADP is thus at a site that retains a catalytic capacity for slow single-site ATP hydrolysis (or synthesis) and is likely the site that participates in cooperative rapid net ATP hydrolysis. During hydrolysis of 50 microM [3H]ATP in the presence of either Mg2+ or Ca2+, the enzyme has a steady-state level of about one bound [3H]ADP per mole of enzyme. Because bound [3H]ATP is also present, the [3H]ADP is regarded as being present on two cooperating catalytic sites. The formation and levels of bound ATP, ADP, and Pi show that reversal of bound ATP hydrolysis can occur with either Ca2+ or Mg2+ present. They do not reveal why no phosphate oxygen exchange accompanies cleavage of low ATP concentrations with Ca2+ in contrast to Mg2+ with the heat-activated enzyme. Phosphate oxygen exchange does occur with either Mg2+ or Ca2+ present when low ATP concentrations are hydrolyzed with the octyl glucoside activated ATPase. Ligand binding properties of Ca2+ at the catalytic site rather than lack of reversible cleavage of bound ATP may underlie lack of oxygen exchange under some conditions.
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PMID:Bound adenosine 5'-triphosphate formation, bound adenosine 5'-diphosphate and inorganic phosphate retention, and inorganic phosphate oxygen exchange by chloroplast adenosinetriphosphatase in the presence of Ca2+ or Mg2+. 287 34

To investigate the behaviour of glycoprotein and glycolipid receptors at the lymphocyte cell surface, a spin label probe has been introduced into either sialic acid or galactose residues on lymphocyte plasma membrane, using specific activation of sugars with periodate or galactose oxidase, followed by reductive amination. The extent of membrane labelling could be controlled by varying the mole ratios of reactants used. Chloroform-methanol extraction of the labelled membranes showed that approximately 17% of the label is bound to glycolipids. A large fraction of the spin label could be released from both sialic acid and galactose-labelled membrane by treatment with pronase, indicating attachment to membrane proteins. Rotational correlation times (tau c) for both labelled sialic acid and galactose residues were in the range 10-13 X 10(-10) sec, indicating a reduction in sugar headgroup mobility at the membrane surface. Isolated lymphocyte membrane glycoproteins spin labelled on galactose residues and reassembled into phospholipid bilayer vesicles showed similar motional characteristics. Prolonged incubation of conc. suspensions of labelled membrane resulted in cleavage of the sialic acid-bound (but not the galactose-bound) label. Binding of several lectins to labelled plasma membrane produced significant immobilization of cell surface oligosaccharides while others had no effect. This differential restriction in oligosaccharide motion following lectin binding appears to be at least partly related to the sugar specificity of the lectin. Binding of wheat germ agglutinin and Ricinus communis agglutinin to sialic acid and galactose-labelled membrane respectively produced a dramatic decrease in oligosaccharide mobility which was reversible on addition of the appropriate sugar inhibitor. The concn dependence of lectin-induced spin label immobilization suggested a cooperative interaction between the lectins and their oligosaccharide receptors. Binding of lectins to the lymphocyte cell surface thus seems to have distinct effects on the dynamic state of glycoproteins and glycolipids within the glycocalyx.
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PMID:Spin labelling of sialic acid and galactose residues on lymphocyte plasma membrane: effects of lectins on oligosaccharide dynamics. 299 54

Efficient delivery of hydrophobic water-insoluble substrates and cofactors to membrane-bound enzymes is a recurring problem which has impeded kinetic analyses. Kinetic analysis of the Escherichia coli sn-1,2-diacylglycerol kinase, an extremely hydrophobic integral membrane protein of 122 residues, was facilitated by the development of a mixed micellar assay. beta-Octyl glucoside micelles quantitatively solubilized diacylglycerol kinase from membranes of strains which overproduced the enzyme up to 250-fold and provided an effective method to disperse and deliver the hydrophobic water-insoluble substrate, sn-1,2-dioleoyglycerol. Diacylglycerol kinase was active in mixed micelles containing octyl glucoside and dioleoyglycerol. Several phospholipids stimulated activity up to 6-fold, suggesting a cofactor function. Activation by phospholipids was not stereospecific and was mimicked partially by fatty acids. Half-maximal activation was observed at 1 mol % cardiolipin, suggesting that a small number of phospholipids are sufficient to activate the enzyme. Activity was dependent on the mole fractions of dioleoylglycerol and phospholipid in the mixed micelles, but independent of micelle number. Several lines of evidence indicated that the transfer of diacylglycerol between micelles was much more rapid than its utilization by the enzyme. Diacylglycerol kinase exhibited Michaelis-Menten kinetics with respect to diacylglycerol and MgATP. A second Mg2+ ion (in addition to MgATP) was required for activity. When Mg2+ was excluded from the assay, Mn2+, Zn2+, Cd2+, and Co2+ supported activity to lesser extents. These data establish a suitable system for in-depth kinetic analysis of the E. coli diacylglycerol kinase and its phospholipid cofactor requirements.
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PMID:sn-1,2-Diacylglycerol kinase of Escherichia coli. Mixed micellar analysis of the phospholipid cofactor requirement and divalent cation dependence. 300 49

The extracellular hemoglobin of Lumbricus terrestris (3900 kDa) consists of at least six different polypeptide chains: I through IV (16-19 kDa), V (31 kDa) and IV (37 kDa) (Vinogradov, S.N., Shlom, J.M., Hall, B.C., Kapp, O.H. and Mizukami, H. (1977) Biochim. Biophys. Acta 492, 136-155). SDS-polyacrylamide gel electrophoresis of the unreduced hemoglobin shows that chains II, III and IV form a disulfide-bonded 50 kDa subunit. This subunit was isolated by gel filtration of the hemoglobin on Sephacryl S-200 (a) at neutral pH in 0.1% SDS and (b) in 0.1 M sodium acetate buffer (pH 4.0); in the latter case it retains heme. The 50 kDa subunit obtained by method (b) was reduced and subjected to chromatofocusing on PBE 94 column: the elution pattern obtained with Polybuffer 74 (pH 4.5) and monitored at 280 nm, consisted of three peaks A, B and C; peaks A and B but not C, had absorbance at 410 nm. SDS-polyacrylamide gel electrophoresis showed that peaks A, B and C corresponded to chains II, IV and III, respectively. Amino acid analyses and N-terminal sequence determinations identified chain II as the whose primary structure had been determined (Garlick, R. and Riggs, A. (1982) J. Biol. Chem. 257, 9005-9015). Carbohydrate analysis of the native hemoglobin shows it to contain 2.0 +/- 0.5% carbohydrate consisting of mannose and N-acetylglucosamine in a mole ratio of about 9:1. The carbohydrate content of the 50 kDa subunit is 1.8 +/- 0.5%; it consists of mannose and N-acetylglucosamine in the same ratio and it appears to be associated with chain IV. Rabbit polyclonal antisera to 50 kDa subunit, prepared by method (a), and to the native hemoglobin were shown to cross-react with the hemoglobin and the 50 kDa subunit, respectively, by immunodiffusion. One of eight mouse monoclonal antibodies directed against the native hemoglobin reacted strongly with the 50 kDa subunit prepared by methods (a) and (b) in an enzyme-linked immunosorbent assay (ELISA). Another monoclonal antibody reacted strongly with the 50 kDa subunit obtained by method (b). Neither of the two hybridomas exhibited a strong reaction with any of the three constituent chains of the 50 kDa subunit. These results suggest that the unusual disulfide-bonded 50 kDa subunit, consisting of three myoglobin-like polypeptide chains of which only two have heme, is an integral part of the native Lumbricus hemoglobin molecule.
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PMID:A disulfide-bonded trimer of myoglobin-like chains is the principal subunit of the extracellular hemoglobin of Lumbricus terrestris. 308 Oct 31

The isolated perfused rat liver preparation was employed to study hepatic disposition of the model drug-carrier conjugate fluorescein-lactosylated albumin (F-LnHSA) with special reference to the influence of the organic anion fluorescein on liver cell specificity of the endocytosed neoglycoprotein. Hepatic clearance of fluoresceinated neoglycoproteins was significantly faster than clearance of radioiodinated neoglycoproteins. Perfusate clearance of F-L7HSA and F-L25HSA could not completely be inhibited by a dose of 10 mg asialoorosomucoid that saturates the hepatocyte receptor-mediated endocytic process. From these data, we inferred an additional hepatic uptake mechanism, competing with the Ashwell-receptor-mediated internalization of galactose-terminated glycoproteins. Clearance experiments with fluoresceinated 125I-human serum albumin in the presence of the polyanionic probe dextran sulfate revealed a nearly complete (approximately 90%) inhibition of hepatic uptake, while also a pronounced effect was obtained with colloidal carbon. These data point to nonparenchymal cell uptake of fluoresceinated protein via interaction with scavenger receptors. In wash-out studies, it was shown that about 25% of ligand sequestrated by sinusoidal liver cells escaped degradation and recycled to the perfusion medium. Our results show that care should be taken in the use of neoglycoproteins as drug carriers to hepatocytes, since a load of only 2 to 3 moles fluorescein per mole neoglycoprotein considerably affects intrahepatic distribution. The relative contribution of nonparenchymal cell uptake by coupling of acidic drugs to the neoglycoproteins is very probably inversely related to the number of exposing galactose groups per molecule neoglycoprotein. This phenomenon of "inversed targeting" could therapeutically both be useful or detrimental, dependent on the spectrum of cell types that should be reached by the drug.
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PMID:Drug targeting to the liver with lactosylated albumins: does the glycoprotein target the drug or is the drug targeting the glycoprotein? 308 97

A simple and convenient technique has been developed for detection of beta-galactosidase from E. coli on nitrocellulose sheets using a mixture of 5-bromoindol-3-yl-beta-D-galactopyranoside and nitro blue tetrazolium, which enables rapid detection of fmole (10(-15) mole) quantities of the enzyme at pH 9.5. The technique has the following advantages: the substrates are stable for a long period; reaction products give non-fading intense blue colour, resolution is extremely good with essentially no diffusion.
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PMID:[Determination of beta-galactosidase activity on nitrocellulose plates using 5-bromo-3-indolyl-beta-D-galactopyranoside and tetrazolium salts]. 310 76

Using potato dextrose agar medium, 40 strains of microorganisms were isolated from leftover fermented corn flour samples involved in outbreaks of food poisoning. All strains produced powerful toxins which caused the same intoxication to mice, dogs, and monkeys as the leftover food samples. On the basis of results obtained from the morphology of this bacteria and its colony, from biochemical tests, and from the G-C mole percentage in DNA, the bacteria was identified as Flavobacterium farinofermentans nov. sp. (Meng, Z. and Wang, D.).
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PMID:Studies on fermented corn flour food poisoning in rural areas of China. II. Isolation and identification of causal microorganisms. 315 54


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