Kimiko Anno

Mucopolysaccharide-degrading enzymes from the liver of the squid, Ommastrephes sloani pacificus. I. Hyaluronidase.

Abstract 1. 1. Liver extracts of the squid, Ommastrephes sloani pacificus , contain mucopolysaccharide-degrading enzymes: hyaluronidase (hyaluronate glycanohydrolase, EC 3.2.1.35), β- N -acetylhexosaminidase (EC 3.2.1.30); β-glucuronidase (EC 3.2.1.31) and chondrosulphatase (EC 3.1.6.4). 2. 2. The squid hyaluronidase which was partially purified by gel chromatography on Sephadex G-150 showed the maximum activity at pH 4.5–5.0 and the temperature of 37°. The activity was completely inhibited by Cu 2+ and Fe 3+ . 3. 3. Hyaluronate was the best substrate of the squid hyaluronidase and several sulphated mucopolysaccharides were also degraded. The digestion products of hyaluronate were identical with those obtained by bovine testicular hyaluronidase digestion, indicating that the squid liver hyaluronidase is an endo- β - N -aacetylhexosaminidase. 4. 4. This seems to be the first example of a testicular type hyaluronidase found in invertebrate tissues.

A modified method for chondrosulfatase assay

The turbidimetric method of Dodgson and Price has been modified for chondrosulfatase assay. The method involves removal of undegraded chondroitin sulfates with cetylpyridinium chloride, modification of the barium chloride-gelatin reagent, and substitution of 0.2N hydrochloric acid for the 3% trichloroacetic acid of the original method. By using this modified method, the presence of chondrosulfatase activity was demonstrated in the liver of the squid, Ommastrephes sloani pacificus.

A new dermatan polysulfate, chondroitin sulfate H, from hagfish notochord

Abstract A novel oversulfated mucopolysaccharide was isolated from the notochord of hagfish, which belongs to the more primitive branch of cyclostomes. It is concluded that the mucopolysaccharide is a new type of dermatan polysulfate consisting mainly of (1 → 4)-α-L-idopyranuronosyl-(1 → 3)-2-acetamido-2-deoxy-β-D-galactopyranosyl 4,6-disulfate unit. The name of chondroitin sulfate H is proposed for this dermatan polysulfate.

Mucopolysaccharides from chicken skin of three age groups.

Abstract Mucopolysaccharides were isolated from the skin of chickens of three age groups: newly hatched, 4 weeks old and 23 months old, and determined by electrophoresis on cellulose acetate strips, Dowex I column chromatography and chondroitinase digestion. Hyaluronic acid and dermatan sulfate were the major components in all age groups; the content of hyaluronic acid decreased and dermatan sulfate increased with age. Small amounts of chondroitin sulfate A, dermatan polysulfate and a heparan sulfate-like substance were also found. The mucopolysaccharide pattern of 4-week-old chicken skin was closer to that of 23-month-old skin than that of newly hatched.

Isolation and characterization of a new disaccharide disulfate: 2-acetamido-2-deoxy-3-O-(2-or 3-O-sulfo-β-D- glucopyranosyluronic acid)-4-O-sulfo-D-galactose

Abstract A new disaccharide disulfate was isolated, in high yield, from a chondroitin polysulfate of king crab cartilage. The structure of this disaccharide was characterized as 2- acetamido -2- deoxy -3-O-(2- or 3-O- sulfo -β- D - glucopyranosyluronic acid )-4-O- sulfo- D -galactose .

A novel dermatan polysulfate from hagfish skin, containing trisulfated disaccharide residues.

Abstract A novel type of dermatan polysulfate was isolated from the skin of the hagfish. Digestion of this mucopolysaccharide with chondroitinase-ABC yielded a trisulfated disaccharide, which was characterized as 2-acetamido-2-deoxy-3- O -(2- or 3- O -sulfo- β -D-gluco-4-enepyranosyluronic acid)-4,6-di- O -sulfo-D-galactose, in addition to 2-acetamido-2-deoxy-3- O -(2- or 3- O -sulfo- β -D-gluco-4-enepyranosyluronic acid)-4- O -sulfo-D-galactose; 2-acetamido-2-deoxy-3- O -( β -D-gluco-4-enepyranosyluronic acid)-4,6- O -disulfo-D-galactose; 2-acetamido-2-deoxy-3- O -( β -D-gluco-4-enepyranosyluronic acid)-4- O -sulfo-D-galactose and -6- O -sulfo-D-galactose. This is the first isolation of a disaccharide trisulfate from dermatantype mucopolysaccharides.

Substrate specificity of chondrosulfatases from Proteus vulgaris for sulfated tetrasaccharides

1. 1. Three novel sulfated tetrasaccharides, III, IV and V were isolated from the digestion products of chondroitin sulfate C, E and K, with testicular hyaluronidase, and were characterized as follows: III,O-(β-D-glucopyranosyluronic acid)-(1→3)-O-2acetamido-2-deoxy-6-O-sulfo-β-D-gala ctopyranosyl-(1→4)-O-(β-D-glucopyranosyluronicacid)-(1→3)-2-acetamido-2-deoxy-4-O-sulfo-D-galactose; IV,O-(β-D-glucopyranosyluronicacid)-(1→3)-O-2-acetamido-2-deoxy-4,6-di-O-sulfo-β-D-galactopyranosyl-(1→4)-O -(β-D-glucopyranosyluronicacid)-(1→3)-2-acetamido-2-deoxy-4-O-sulfo-D-galactose; V,O-(2-or3-O-sulfo-β-D-glucopyranosyluronic acid)-(1→3)-O-2-acetamido-2-deoxy-4-O-sulfo-β-D-galactopyranos yl-(1→4)-O-(β-D-glucopyranosyluronicacid)-(1→3)-2-acetamido-2-deoxy-4-O-sulfo-D-galactose. 2. 2. Effects of chondro-4- and -6-sulfatases of Proteus vulgaris were studied on III, IV, V, in addition to the tetrasaccharides I, O - (β - D - glucopyranosyluronic acid)-(1→3)-O-2-acetamido-2-deoxy-4-O- sulfo-β-D-galactopyranosyl-(1→4)-O-(β-D-glucopyranosyluronicacid)-(1→3)-2-acetamido-2-deoxy-4-O-sulfo-D-galactose and II, O-(β-D-glucopyranosyluronic acid)-(1→3)-O-2-acetamido-2-deoxy-6-O-sulfo-β-D-galactopyranosyl-(1→4)-O (β-D-glucopyranosyluronicacid)-(1→3)-2-acetamido-2-deoxy-6-O-sulfo-D-galactose. 3. 3. The chondro-4-sulfatase attacked preferentially the ester sulfate on the C-4 of the N-acetylgalactosamine residue in the reducing end of the tetrasaccharide I, III, IV, V, regardless of the positions of the ester sulfates in the non-reducing disaccharide moieties, and then gradually attacked the ester sulfate on the C-4 of the other side in I and V. On the other hand, chondro-6-sulfatase scarcely affected the C-6 ester sulfate of sulfated tetrasaccharides.

A chitin sulfate-like polysaccharide from the test of the tunicate Halocynthia roretzi

Abstract Considerable amounts of water-soluble polysaccharides were found in the test of the tunicate Halocynthia roretzi . After fractionation with ethanol, a chitin sulfate-like polysaccharide was isolated. From the results of chemical analysis, optical rotation, infrared spectrum and alkaline treatment, it is suggested that the predominant structure for this polysaccharide is (1→4)-2-acetamido-2-deoxy-6- O -sulfo- β -D-glucopyranan.

Isolation of heparan sulfate from chicken skin

Abstract Two heparan sulfate fractions (1.0 and 1.25 M heparan sulfate) were isolated from chicken skin by Dowex 1 column chromatography and testicular hyaluronidase digestion followed by Sephadex G-50 gel chromatography. The yields of 1.0 and 1.25 M heparan sulfate were 2.1 and 1.3% of the total mucopolysaccharides, respectively. From the results of chemical analysis, infrared spectroscopy and electrophoresis, these two fractions were found to be essentially identical, except for their sulfate contents. This seems to be the first example of heparan sulfate isolated from non-mammalian skin.

A comparison of glucosamine and galactosamine content of cartilage from various sources

Abstract 1. 1. A comparative study was made, by using ion-exchange chromatography, of the glucosamine and galactosamine contents of various cartilages obtained from the following animals: whale, cattle, ray, shark and squid. 2. 2. The cartilages analyzed here all contain glucosamine as well as galactosamine, indicating the presence of glucosamine-containing mucopolysaccharides in addition to chondroitin sulfate. 3. 3. The glucosamine content varies with the origin; it is less in elasmobranchs than in mammals, and least in the cephalopod. 4. 4. The relative proportions of glucosamine and galactosamine in adult cartilage appear to depend on the species and the region of the animal body; a similar ratio was found in the same region of animals belonging to the same zoological class, while different ratios were obtained from different regions even in the same animal body. 5. 5. The glucosamine contents in the whale and ray fetus were much smaller than those in the respective adult animals.