scholarly journals Skeletal keratan sulphate chains isolated from bovine intervertebral disc may terminate in α(2----6)-linked N-acetylneuraminic acid

1992 ◽  
Vol 282 (1) ◽  
pp. 267-271 ◽  
Author(s):  
J M Dickenson ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Intervertebral Disc ◽  
Anion Exchange ◽  
Gel Permeation Chromatography ◽  
Intervertebral Discs ◽  
Borohydride Reduction ◽  
Anion Exchange Column ◽  
Keratan Sulphate ◽  
Acetylneuraminic Acid ◽  
Sialidase Inhibitor ◽  
Gel Permeation

Peptido-keratan sulphate fragments were isolated from the nucleus pulposus of bovine intervertebral discs (2-year-old animals) after digestion with chondroitin ABC lyase followed by digestion with diphenylcarbamoyl chloride-treated trypsin of A1D1 proteoglycans and gel-permeation chromatography on Sepharose CL-6B. The peptido-keratan sulphate fragments were subjected to alkaline borohydride reduction. The reduced chains were treated with keratanase in the presence of the sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid, and the digest was subjected to alkaline borohydride reduction. This produced oligosaccharides with galactitol at their reducing ends. This reduced digest was chromatographed on a Nucleosil 5 SB anion-exchange column and individual oligosaccharides were isolated. One of these was shown by 600 MHz 1H-n.m.r. spectroscopy to have the following structure: NeuAc alpha 2-6Gal beta 1-4GlcNAc(6-SO4)beta 1-3Gal-ol The structure of this oligosaccharide shows that keratan sulphate chains from bovine intervertebral disc have non-reducing termini with N-acetylneuraminic acid linked alpha(2----6) as well as alpha(2----3) to an unsulphated galactose.

scholarly journals A non-reducing terminal fragment from tracheal cartilage keratan sulphate chains contains α(2-3)-linked N-acetylneuraminic acid

1991 ◽  
Vol 278 (3) ◽  
pp. 779-785 ◽  
Author(s):  
J M Dickenson ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Anion Exchange ◽  
Chondroitin Sulphate ◽  
Tracheal Ring ◽  
Exchange Chromatography ◽  
Borohydride Reduction ◽  
Anion Exchange Column ◽  
Keratan Sulphate ◽  
Sialidase Activity ◽  
Acetylneuraminic Acid ◽  
Femoral Head Cartilage

Keratan sulphate chains were isolated from bovine tracheal ring cartilage (15-18-month-old animals) after papain digestion of the tissue followed by ethanol fractionation, chondroitinase ABC digestion and alkaline borohydride reduction. The keratan sulphate chains were further purified by anion-exchange chromatography on a Pharmacia Mono-Q column in order to remove any contaminating chondroitin sulphate and O-linked oligosaccharides. The chains were then treated with keratanase and the digest was subjected to alkaline borohydride reduction, producing oligosaccharides with galactitol at their reducing ends. The reduced digest was chromatographed on a Nucleosil 5 SB anion-exchange column and individual oligosaccharides were isolated. One of these, oligosaccharide (I), was shown by 500 MHz 1H-n.m.r. spectroscopy to have the following structure: NeuAc alpha 2-3Gal beta 1-4GlcNAc(6SO4) beta 1-3Gal-ol (I) The structure of this oligosaccharide shows that keratan sulphate chains from bovine tracheal ring cartilage may be terminated with N-acetylneuraminic acid linked alpha (2-3) to an unsulphated galactose. Keratan sulphate chains were also isolated from bovine femoral head cartilage (15-18-month-old animals) using an identical protocol, but with keratanase which was subsequently shown to have sialidase activity. This yielded oligosaccharide (II), the unsialyated version of (I): Gal beta 1-4GlcNAc(6SO4) beta 1-3Gal-ol (II).

scholarly journals Two linkage-region fragments isolated from skeletal keratan sulphate contain a sulphated N-acetylglucosamine residue

1990 ◽  
Vol 269 (1) ◽  
pp. 55-59 ◽  
Author(s):  
J M Dickenson ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Gel Permeation Chromatography ◽  
Anion Exchange Chromatography ◽  
Ester Group ◽  
Intervertebral Discs ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Keratan Sulphate ◽  
Linkage Region ◽  
Gel Permeation ◽  
Electron Micro Probe Analysis

Peptido-keratan sulphate fragments were isolated from the nucleus pulposus of bovine intervertebral discs (6-year-old animals) after chondroitin ABC lyase digestion followed by digestion of A1D1 proteoglycans by diphenylcarbamoyl chloride-treated trypsin and gel-permeation chromatography on Sepharose CL-6B. Treatment of these peptido-keratan sulphate fragments with alkaline NaB3H4 yielded keratan sulphate chains with [3H]galactosaminitol end-labels, and these chains were further purified by gel-permeation chromatography on Sephadex G-50 and ion-exchange chromatography on a Pharmacia Mono-Q column in order to exclude any contamination with O-linked oligosaccharides. The chains were then treated with keratanase, and the digest was chromatographed on a Bio-Gel P-4 column followed by anion-exchange chromatography on a Nucleosil 5 SB column. Two oligosaccharides, each representing 18% of the recovered radiolabel, were examined by 500 MHz 1H-n.m.r. spectroscopy, and shown to have the following structures: [formula: see text] The structure of oligosaccharide (I) confirms the N-acetylneuraminylgalactose substitution at position 3 of N-acetylgalactosamine in the keratan sulphate-protein linkage region found by Hopwood & Robinson [(1974) Biochem. J. 141, 57-69] but additionally shows the presence of a 6-sulphated N-acetylglucosamine. Electron micro-probe analysis specifically confirmed the presence of sulphur in this sample. This sulphate ester group differentiates the keratan sulphate linkage region from similar structures derived from O-linked oligosaccharides [Lohmander, De Luca, Nilsson, Hascall, Caputo, Kimura & Heinegård (1980) J. Biol. Chem. 255, 6084-6091].

scholarly journals Fucose content of keratan sulphates from bovine articular cartilage

1991 ◽  
Vol 273 (2) ◽  
pp. 307-310 ◽  
Author(s):  
G H Tai ◽  
G M Brown ◽  
H G Morris ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Molecular Weight ◽  
Articular Cartilage ◽  
Molecular Size ◽  
Gel Permeation Chromatography ◽  
Repeat Sequence ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Gel Permeation ◽  
Fucose Content ◽  
Galactose Content

Alkaline-borohydride-reduced keratan sulphate chains were isolated from bovine articular cartilage (6-8-year-old animals). Nine keratan sulphate fractions of increasing molecular weight were prepared by gel-permeation chromatography on a calibrated column of TSK 30 XL. The samples were analysed for fucose and galactose contents (% by wt. of keratan sulphate) and fucose/galactose ratio. The fucose content increased with molecular size, but the galactose content remained constant. It was concluded that the alpha(1→3)-linked fucose [Thornton, Morris, Cockin, Huckerby, Nieduszynski, Carlstedt, Hardingham & Ratcliffe (1989) Biochem. J. 260, 277-282] was located within the poly-N-acetyl-lactosamine repeat sequence of articular-cartilage keratan sulphate.

Analysis of 188Re-EDTMP complexes by HPLC and ultrafiltration

2004 ◽  
Vol 92 (4-6) ◽  
Author(s):  
Kazuyuki Hashimoto ◽  
Hiromitsu Matsuoka
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Anion Exchange ◽  
High Performance ◽  
Gel Permeation Chromatography ◽  
Reversed Phase ◽  
Ion Pair ◽  
Gel Permeation

AbstractHigh performance liquid chromatography (anion-exchange, reversed-phase ion-pair and gel permeation chromatography) and ultrafiltration have been employed to analyze

SHORT COMMUNICATIONS Preparative-scale fractionation of bovine, caprine and ovine whey proteins by gel permeation chromatography

1997 ◽  
Vol 64 (3) ◽  
pp. 459-464 ◽  
Author(s):  
XAVIER FELIPE ◽  
ANDREW J. R. LAW
Keyword(s):  
Whey Protein ◽  
Anion Exchange ◽  
Whey Proteins ◽  
Gel Permeation Chromatography ◽  
Reversed Phase ◽  
Complete Separation ◽  
Protein Fractions ◽  
Rapid Preparation ◽  
Preparative Scale ◽  
Gel Permeation

The whey proteins of the cow, goat and sheep have previously been fractionated on an analytical scale by reversed-phase HPLC (De Frutos et al. 1992), anion-exchange FPLC (Andrews et al. 1985; Manji et al. 1985; Laezza et al. 1991) and gel permeation FPLC (Andrews et al. 1985; Hill & Kakuda, 1990). Anion-exchange and gel permeation FPLC can readily be scaled up for laboratory preparation of whey protein fractions. There is some indication, however, that anion-exchange FPLC does not give complete separation of β-lactoglobulin and α-lactalbumin from the other minor whey protein fractions (Girardet et al. 1989).In previous work it has been shown that gel permeation FPLC gives a satisfactory fractionation of the whey proteins of the cow (Law et al. 1993), goat (Law & Brown, 1994) and sheep (Law, 1995). In this paper we describe a scaled-up method of gel permeation that can be used for fairly rapid preparation or purification of four main whey protein fractions from the milks of these species.

scholarly journals A sub-population of keratan sulphates derived from bovine articular cartilage is capped with α(2–6)-linked N-acetylneuraminic acid residues. Affinity chromatography using immobilized Sambucus nigra lectin and characterization using 1H n.m.r. spectroscopy

1992 ◽  
Vol 286 (1) ◽  
pp. 231-234 ◽  
Author(s):  
G H Tai ◽  
H G Morris ◽  
G M Brown ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Affinity Chromatography ◽  
Gel Permeation Chromatography ◽  
Similar Proportion ◽  
Lectin Affinity Chromatography ◽  
Sambucus Nigra ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Acetylneuraminic Acid ◽  
Sambucus Nigra Agglutinin ◽  
Femoral Head Cartilage

Alkaline borohydride-reduced keratan sulphate (KS) chains derived from bovine femoral head cartilage were fractionated by lectin affinity chromatography with Sambucus nigra agglutinin (SNA) into binding and non-binding populations. Analysis of the SNA-binding and non-binding KS chains using 600 MHz 1H n.m.r. spectroscopy showed that the former population contained alpha(2-6)-N-acetylneuraminic acid residues and the latter contained primarily alpha(2-3)-N-acetylneuraminic acid residues as chain terminators. Both populations contained a similar proportion of alpha(2-3)-N-acetylneuraminic acid residues within their protein-linkage regions, and similar sulphation and fucosylation levels. Analysis of these two fractions by gel-permeation chromatography (g.p.c.) on a TSK-30 XL column showed them to have the same size distributions. It was concluded from the n.m.r. spectra and g.p.c. data that the populations differed primarily in the mode of linkage of the chain-terminating sialic acids.

scholarly journals Chemical structure and biological properties of a polysaccharide isolated from Pleurotus sajor-caju

RSC Advances ◽  
2019 ◽  
Vol 9 (35) ◽  
pp. 20472-20482 ◽  
Author(s):  
Palaniappan Seedevi ◽  
Abirami Ramu Ganesan ◽  
Kannan Mohan ◽  
Vasantharaja Raguraman ◽  
Murugesan Sivakumar ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Column Chromatography ◽  
Chemical Structure ◽  
Biological Properties ◽  
Anion Exchange Column ◽  
Exchange Column ◽  
Gel Permeation ◽  
Pleurotus Sajor Caju

Herein, a polysaccharide obtained from Pleurotus sajor-caju was fractionated via anion-exchange column chromatography and purified using gel permeation column chromatography.

scholarly journals The structure and composition of cartilage keratan sulphate

1974 ◽  
Vol 141 (2) ◽  
pp. 517-526 ◽  
Author(s):  
John J. Hopwood ◽  
H. Clem Robinson
Keyword(s):  
Intervertebral Disc ◽  
Nasal Septum ◽  
Alkali Treatment ◽  
Neuraminic Acid ◽  
Keratan Sulphate ◽  
Molecular Parameters ◽  
Acetylneuraminic Acid ◽  
Different Types

Keratan sulphate was isolated from bovine intervertebral disc and bovine nasal septum after hydrolysis with proteinases and treatment with dilute alkali. Each preparation was found to contain, per keratan sulphate chain: (a) 1 residue of mannose; (b) 3 residues of N-acetylneuraminic acid (2 residues after alkali treatment); (c) 1 residue of N-acetylgalactosamine (lost after alkali treatment); (d) 1 residue or less of fucose. N-Acetyl-neuraminic acid residues were at non-reducing termini and were bonded to keratan sulphate through galactose residues. Evidence is presented for two different types of linkage between skeletal keratan sulphate and protein. Consideration of molecular parameters and compositions leads to a proposed structure for keratan sulphate–protein as found in skeletal proteoglycans.

scholarly journals The isolation and properties of a second glycoprotein (LGP-II) from the articular lubricating fraction from bovine synovial fluid

1979 ◽  
Vol 179 (3) ◽  
pp. 465-471 ◽  
Author(s):  
D A Swann ◽  
G Mintz
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Synovial Fluid ◽  
Bovine Serum ◽  
Gel Permeation Chromatography ◽  
Sedimentation Equilibrium ◽  
Major Constituent ◽  
Precipitin Line ◽  
Acetylneuraminic Acid ◽  
Gel Permeation

A high-molecular-weight glycoprotein (LGP-I) was shown [Swann, Sotman, Dixon & Brooks (1977) Biochem. J. 161, 473–485] to be the major constituent in the articular lubricating fraction from bovine synovial fluid. In addition to the LGP-I component, a second glycoprotein (LGP-II) was also present. After fractionation of bovine synovial fluid by sequential sedimentation in CsCl density gradients, the LGP-I and LGP-II components were separated by gel-permeation chromatography. The LGP-II component was then purified by chromatography on DEAE Bio-Gel A and Bio-Gel P-150. The molecular weight of the LGP-II component was 48,800 calculated from sedimentation-equilibrium measurements. Amino acids represented 53% (w/w) and carbohydrate constituents 36% (w/w) of the molecule. Glutamic acid and lysine (144 and 100 residues/1000 residues) were the major amino acids. Glucosamine, mannose, galactose and N-acetylneuraminic acid [representing 8.0, 6.6, 9.5 and 11.9% (w/w) respectively] were the only carbohydrate constituents detected. Immunodiffusion analysis showed that LGP-II component did not form a detectable precipitin line with antiserum to bovine serum. It appears likely, therefore, that this glycoprotein is synthesized by the joint tissues and is not derived from serum.

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