scholarly journals Preliminary characterization of the oligosaccharide component of arylsulfatase B from human placenta.

1995 ◽  
Vol 42 (1) ◽  
pp. 45-50
Author(s):  
P Laidler ◽  
M Gałka-Walczak ◽  
A Lityńska
Keyword(s):  
Sialic Acid ◽  
Human Placenta ◽  
Carbohydrate Component ◽  
Lectin Affinity Chromatography ◽  
Complex Type ◽  
Total Carbohydrate ◽  
Chain Analysis ◽  
Glycan Chain ◽  
Arylsulfatase B

Isoelectric focusing of homogenous arylsulfatase B from human placenta pointed to the presence of enzymatically active and inactive forms of high pI (pH 9-8) and of lower pI (pH 6.5-5.5). Glycan chain analysis performed with the use of a Glycan Differentiation Kit showed that basic forms of arylsulfatase B from human placenta contained mostly high mannose/hybrid type glycans, with 6-O-L-fucose bound to the innermost N-acetylglucosamine residue, whereas acidic forms of the enzyme contained complex type glycans containing fucose and sialic acid. However, the latter forms constitute a small percentage of the total carbohydrate component. Lectin affinity chromatography of the native enzyme confirmed the presence of a core fucose and a sialic acid.

scholarly journals Characterization of the oligosaccharide component of arylsulfatase B from rat liver.

1997 ◽  
Vol 44 (2) ◽  
pp. 181-190 ◽  
Author(s):  
M Przybyło ◽  
A Lityńska
Keyword(s):  
Rat Liver ◽  
Gel Permeation Chromatography ◽  
Complex Type ◽  
Relative Contribution ◽  
Chain Analysis ◽  
Terminal Galactose ◽  
Glycan Chain ◽  
Arylsulfatase B ◽  
High Mannose

A glycan chain analysis of the total oligosaccharide pool derived from rat liver arylsulfatase B was carried out by. P4 Gel Permeation Chromatography and sequential exoglycosidase digestion. It was found that 71% of rat liver arylsulfatase B oligosaccharides were sialylated. The relative contribution of particular structures in the total glycan pool was as follows: sialylated biantennary complex type glycans with terminal galactose--65%, high-mannose type glycans--15%, biantennary complex type glycans with core fucose and terminal N-acetylglucosamine--5%, O-linked oligosaccharides--3.5%.

scholarly journals Purification and enzymatic characterization of CMP-sialic acid: beta-galactosyl1----3-N-acetylgalactosaminide alpha 2----3-sialyltransferase from human placenta.

1985 ◽  
Vol 260 (8) ◽  
pp. 4941-4951
Author(s):  
D H Joziasse ◽  
M L Bergh ◽  
H G ter Hart ◽  
P L Koppen ◽  
G J Hooghwinkel ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Human Placenta ◽  
Enzymatic Characterization

scholarly journals Expanding the Reaction Space of Linkage-Specific Sialic Acid Derivatization

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3617 ◽  
Author(s):  
Tamas Pongracz ◽  
Manfred Wuhrer ◽  
Noortje de Haan
Keyword(s):  
Sialic Acid ◽  
Sialic Acids ◽  
Systematic Evaluation ◽  
Complex Type ◽  
Established Method ◽  
Reaction Space ◽  
Byproduct Formation ◽  
High Degree

The human glycome is characterized by a high degree of sialylation, affecting, amongst others, cell–cell interactions and protein half-life. An established method for the linkage isomer-specific characterization of N-glycan sialylation is based on the linkage-specific derivatization of sialylated glycoconjugates, inducing ethyl esterification of α2,6-linked sialic acids and lactonization of α2,3-linked sialic acids. While the carboxylic acid activator and nucleophile used in this reaction received extensive investigation, the role of the catalyst was never thoroughly explored. A frequently used catalyst for the linkage-specific esterification of sialic acids is 1-hydroxybenzotriazole (HOBt). Here, a systematic evaluation was performed of five HOBt alternatives in combination with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) in ethanol for the linkage-specific derivatization of sialic acids. Derivatized glycans were analyzed by MALDI-TOF-MS and the catalyst performance was evaluated based on the completeness of the reactions and the linkage-specificity obtained. The use of both 6-Cl-HOBt and 6-CF3-HOBt resulted in high linkage-specificity and minimal byproduct formation, similar to the benchmark method using HOBt. Performing the reaction with these catalysts at neutral or acidic pH showed comparable efficiencies on both sialyllactose and complex-type N-glycans. The reported investigations resulted in an expansion of the reaction space for linkage-specific sialic acid derivatization.

Characterization of the oligosaccharides of liver Z variant α1-antitrypsin

1980 ◽  
Vol 58 (8) ◽  
pp. 644-648 ◽  
Author(s):  
Albert Hercz ◽  
Noam Harpaz
Keyword(s):  
Endoplasmic Reticulum ◽  
Sialic Acid ◽  
Sindbis Virus ◽  
Smooth Endoplasmic Reticulum ◽  
Gel Permeation Chromatography ◽  
New Product ◽  
Complex Type ◽  
Homogeneous Product ◽  
Normal Individuals

The plasma α1-antitrypsin (AT) of normal individuals has been reported to have four oligosaccharides per molecule; these oligosaccharides are all of the complex type, containing only three D-mannose (Man) residues per oligosaccharide as well as several residues of galactose, sialic acid, and N-acetyl-D-glucosamine (GlcNAc) (Chan, S. K., Rees, D. C., Li, S.-C. &Li, Y.-T. (1976) J. Biol. Chem. 251, 471–476; Hodges, L. C., Laine, R. &Chan, S. K. (1979) J. Biol. Chem. 254, 8208–8212). Analysis of the carbohydrates of purified liver Z variant AT was consistent with the presence of three moles of Man-rich oligosaccharide per mole protein with an average of seven Man and two GlcNAc residues per oligosaccharide and a complete absence of galactose and sialic acid (Hercz, A., Katona, E., Cutz, E., Wilson, J. R., &Barton, M. (1978) Science 201, 1229–1232). We have now carried out a more definitive structural analysis of the oligosaccharides of liver Z variant. Glycopeptides were prepared from purified liver Z variant by digestion with pronase. Treatment of a sample of the glycopeptides with endo-β-N-acetylglycosaminidase CII (endo CII) resulted in the formation of three oligosaccharides. These were identified by gel permeation chromatography on a Biogel P-4 column, calibrated with a series of ovalbumin and Sindbis virus oligosaccharides, as GlcNAcMan5, GlcNAcMan6, and GlcNAcMan7. Treatment of a sample of the glycopeptides with endo-β-N-acetylglycosaminidase D gave rise only to a single oligosaccharide product, GlcNAcMan5. Upon incubation of the glycopeptides with UDP[U-14C]GlcNAc in the presence of purified bovine colostrum UDPGlcNAc:α-D-mannoside α-2-N-acetylglucosaminyltransferase I, AsnGlcNAc2Man5 quantitatively disappeared and a new product consistent with the structure AsnGlcNAc2Man5[14C]GlcNAc was formed. Incubation of the oligosaccharides, formed with endo CII, with α-mannosidase gave rise to a 90% homogeneous product with the probable structure of Manβ1 → 4GlcNAc. These findings suggest that the precursors of the complex oligosaccharides of the secretory glycoprotein, plasma AT, are Man-rich oligosaccharides. It appears also that the glycosidases which process the precursor to the AsnGlcNAc2Man5 stage are present and active in the rough endoplasmic reticulum of Pi (protease inhibitor) ZZ individuals; however, for an as yet undetermined reason, most of the Z variant AT appears unable to migrate at a normal rate to the smooth endoplasmic reticulum and Golgi apparatus for eventual secretion from the liver.

scholarly journals Structural characterization of the carbohydrates of the rat ovarian luteinizing hormone/chorionic gonadotropin receptor

1994 ◽  
Vol 298 (2) ◽  
pp. 361-366 ◽  
Author(s):  
U E Petäjä-Repo
Keyword(s):  
Sialic Acid ◽  
Luteinizing Hormone ◽  
Electrophoretic Mobility ◽  
Chorionic Gonadotropin ◽  
Structural Characterization ◽  
Complex Type ◽  
Receptor Molecule ◽  
Gonadotropin Receptor ◽  
Sds Page

The numbers and types of oligosaccharide present on the rat ovarian luteinizing hormone (LH)/chorionic gonadotropin (CG) receptor were determined by treating radiolabelled purified receptors with glycosidases and examining the changes in electrophoretic mobility and number of radiolabelled bands on SDS/PAGE. The purified receptor was also transferred to nitrocellulose after SDS/PAGE and probed with digoxigenin-labelled lectins. The following conclusions were drawn: (1) the rat ovarian LH/CG receptor contains at least two complex-type N-linked oligosaccharide chains, of which one is biantennary and the rest multiantennary. (2) The N-linked chains terminate in either unsubstituted galactose or sialic acid linked alpha 2-3 or alpha 2-6 to the penultimate galactose. (3) The N-linked oligosaccharides also contain internal poly(N-acetyl-lactosamine) sequences and fucose-linked alpha 1-6 to the proximal N-acetylglucosamine. (4) No O-linked carbohydrate moieties are present on the receptor molecule.

scholarly journals Isolation and characterization of methionine synthetase from human placenta.

1985 ◽  
Vol 260 (25) ◽  
pp. 13656-13665 ◽  
Author(s):  
C S Utley ◽  
P D Marcell ◽  
R H Allen ◽  
A C Antony ◽  
J F Kolhouse
Keyword(s):  
Human Placenta ◽  
Isolation And Characterization
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