The Structure of Keratosulfate of Bovine Cornea<sup>1</sup>

Hirano, Shigehiro; Hoffman, Philip; Meyer, Karl

doi:10.1021/jo01070a069

Cited by 74 publications

(11 citation statements)

References 2 publications

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“…Two methods of desulphation were compared. For mild acid-catalysed desulphation (Hirano et al, 1961), dried keratan sulphate of pig nucleus pulposus (20mg) was shaken with anhydrous methanol (2ml) in a tube with Teflon-lined screw cap for 3 h at 24°C. Dry AG 50-X8 resin (H+ form, 200mg) and dry NaCl (40mg) were added, and the heterogeneous mixture was shaken for 48 h at the same temperature.…”

Section: Desulphationmentioning

confidence: 99%

The structure of keratan sulphates from various sources

Choi

Meyer

1975

Biochemical Journal

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Quantitative structural comparisons were made between keratan sulphates isolated from various sources, namely pig nucleus pulposus, bovine cornea, and the costal cartilages of children, a young adult with Marfan syndrome and of old human autopsies. In human costal cartilage the amount of keratan sulphate increases markedly with age, although total mucopolysaccharide decreases to some extent, concomitant with a decrease in chondroitin 4-sulphate and an increase in chondroitin 6-sulphate. Comparison of molecular weights estimated by gel chromatography with those calculated from the molar ratio of galactose to mannose indicates that keratan sulphates of human costal cartilages of children and of a young adult with Marfan syndrome, and of pig nucleus pulposus, contain one mannose residue per chain, whereas keratan sulphates of old human costal cartilage and of bovine cornea contain one to two, and two, per chain respectively. After mild acid-catalysed desulphation of pig nucleus pulposus keratan sulphate, approx. 12% of the mucopolysaccharide aggregates irreversibly once the water is removed from the polysaccharide. The following conclusions have been drawn from a methylation analysis of keratan sulphates of various sources, aided by g.l.c.-mass spectrometry. (1) Fucose and N-acetylneuraminic acid are non-reducing terminal residues and the sialic acid is linked to the 3-position of galactose residues. (2) Pig nucleus pulposus keratan sulphate has approximately 4 non-reducing terminal groups per molecule and appears to be slightly less branched than the costal-cartilage keratan sulphate of children. The branching in human costal-cartilage keratan sulphates decreases with age. Bovine corneal keratan sulphate appears to be unbranched. (3) Mannose residues are linked by 3 different substituents in human costal-cartilage and bovine corneal keratan sulphates, and by two different substituents in pig nucleus pulposus keratan sulphate. (4) The sulphate ester groups are all on the 6-position of N-acetyl-glucosamine and galactose residues. The degree of sulphation increases with age in costal keratan sulphates with the increase mainly of the galactose 6-sulphate residues.

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Section: Desulphationmentioning

confidence: 99%

The structure of keratan sulphates from various sources

Choi

Meyer

1975

Biochemical Journal

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“…Compounds were identified by gas-liquid chromatography retention times and mass spectra. 3 Brought to you by | Purdue Univers Authenticated Download Date | 5/31/15 11:0 Table 4. Glucosamine, galactosamine and amino acid values of the Fractions Con A-l, Con A-2, Con A-3 and EKS.…”

Section: Resultsmentioning

confidence: 99%

Studies on the Characterization of the Linkage-Region between Polysaccharide Chain and Core Protein in Bovine Corneal Proteokeratan Sulfate

Keller¹,

Stein²,

Stuhlsatz³

et al. 1981

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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2,3,4-tri-O-methylfucitol; 2,3,4,6-tetra-O-methyl-1) A new method of enrichment of the linkagemannitol; 3,4,6-tri-O-methylmannitol; 2,4-di-oregion in corneal proteokeratan sulfate is demethylmannitol; 2,3,4,6-tetra-O-methylgalactiscribed, which consists of desulfation of peptidotol; 2,4,6-tri-<9-methylgalactitol; 2,4-di-O-methylkeratan sulfate, followed by chromatography on galactitol. Con A-Sepharose 4B and enzymatic degradation3) The resylts int to the ence of a bnmched with 0-D-galactosidase and 0-TV-acetyl-D-glucosHnkage region in the proteokera tan sulfate ammidase.molecule with one mannose as the branching 2) After permethylation, hydrolysis, reduction point ^ two mannose residues as the starti with sodium borohydrid and acetylation gas int of two disaccharide chains . chromatography/mass spectrometry analyses were performed. The followings products could be detected as their peracetates: Untersuchungen zur Charakterisierung der Bindungsregion zwischen der Polysaccharidkette und dem Core-Protein im Proteokeratansulfat der Rinder-CorneaZusammenfassung:Cornea beschrieben, welche in einer Desulfatie-1) Es wird eine neue Methode zur Anreicherung rung des Peptidokeratansulfats und anschließen-der Bindungsregion im Proteokeratansulfat der den Chromatographie an Con A-Sepharose 4BEnzymes:

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“…Our laboratory has been engaged for a considerable time in a search for an analogous reaction of hexosaminyl moieties in mucopolysaccharides substituted by sulfate in the 6-position and containing a free hydroxyl group in the 3-position. This condition is fulfilled in the keratosulfates (3)(4)(5), heparin (6), and, presumably, heparitin sulfate (7,8). In this communication, the formation of 3,6-anhydroglucosamine by displacement of sulfate in polymers is described for the first time.…”

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confidence: 78%

3,6-Anhydroglucosaminyl Formation in Mucopolysaccharides

Sampson

Meyer

1971

Proc. Natl. Acad. Sci. U.S.A.

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Treatment of keratosulfates with alkali leads to the formation of 3,6-anhydroglucosaminyl groups in high yield. A similar reaction takes place with heparin and heparitin sulfate, though to a lesser extent. The anhydrosugar was isolated and compared with synthetic 3,6-anhydroglucosamine. The isolated compounds were identical in ion exchange chromatography in the amino acid analyzer and on Dowex 50 H+ eluted with dilute HCl. The spectra of the isolated and synthesized anhydrosugar were identical in infrared and nuclear magnetic resonance. Derivatized compounds were identical in gas-liquid chromatography and mass spectroscopy. The peracetylated compound isolated from keratosulfate gave the nuclear magnetic reasonance and fragmentation pattern of the postulated compound. The reaction is expected to be useful in structural studies of hexosamine-containing polymers.The sulfate ester groups of a number of polysaccharides substituted in the 6-position of a galactosyl moiety have been cleaved in an SN2 displacement by alkoxide ion with the concomitant formation of 3,6-anhydrosugar (1, 2). Our laboratory has been engaged for a considerable time in a search for an analogous reaction of hexosaminyl moieties in mucopolysaccharides substituted by sulfate in the 6-position and containing a free hydroxyl group in the 3-position. This condition is fulfilled in the keratosulfates (3-5), heparin (6), and, presumably, heparitin sulfate (7,8). In this communication, the formation of 3,6-anhydroglucosamine by displacement of sulfate in polymers is described for the first time. The reaction will be useful in the fragmentation of these compounds by mild acid hydrolysis. 3,6-anhydro-D-glucosamine was shown by its typical appearance in the amino-acid analyser, by its elution with dilute acid from Dowex 50, by its infrared (IR) and nuclear magnetic resonance (nmr) spectrum, by gas-liquid chromatography (GLC), and by mass spectroscopy. MATERIALSCorneal (KSI) and cartilaginous (KSII) keratosulfates were isolated from bovine cornea and human costal cartilage, respectively, after papain and Pronase digestion, followed by alcohol fractionation. The fractions were further digested with testicular hyaluronidase and chromatographed on Bio-Gel P-100 columns (9). Heparin (119 units/mg) was a gift of the Vitrum Co., Stockholm. Heparitin sulfate, a gift of the Up- (11), and acidified to pH 1-2; the bulk of the borate was removed by distillation from methanol. KSI and KSII were then chromatographed on Bio-Gel P-100 and eluted with 0.5 N NaCl. The product derived from heparin was acidified to pH 4 with glacial acetic acid and eluted from Bio-Gel P-2 with 10% ethanol. The product of heparitin sulfate was acidified to pH 2 with HCl and eluted from Bio-Gel P-2 with 0.5 N NaCl, and the concentrated solution was precipitated with three volumes of ethanol.The displacement reaction of the BH4-reduced compound was performed by heating the compound, at a concentration of 10 mg/ml in 1 N NaOH containing 20 mg/ml of NaBH4, for 7 hr at 800C and reisolatio...

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The Structure of Keratosulfate of Bovine Cornea¹

Cited by 74 publications

References 2 publications

The structure of keratan sulphates from various sources

The structure of keratan sulphates from various sources

Studies on the Characterization of the Linkage-Region between Polysaccharide Chain and Core Protein in Bovine Corneal Proteokeratan Sulfate

3,6-Anhydroglucosaminyl Formation in Mucopolysaccharides

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The Structure of Keratosulfate of Bovine Cornea1

Cited by 74 publications

References 2 publications

The structure of keratan sulphates from various sources

The structure of keratan sulphates from various sources

Studies on the Characterization of the Linkage-Region between Polysaccharide Chain and Core Protein in Bovine Corneal Proteokeratan Sulfate

3,6-Anhydroglucosaminyl Formation in Mucopolysaccharides

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The Structure of Keratosulfate of Bovine Cornea¹