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Y T Li

Publications and source records attributed to Y T Li.

At least 163 records · Page 9Linked to original sources
PubMed1983

The use of mannan-Sepharose 4B affinity chromatography for the purification of endo-beta-N-acetylglucosaminidase from Bacillus alvei.

In order to facilitate the isolation of endo-beta-N-acetylglucosaminidase for the structural analysis of glycoconjugates, we have isolated a strain of Bacillus alvei which produces a high level of endo-beta-N-acetylglucosaminidase. We have also devised a simple procedure for the purification of endo-beta-N-acetylglucosaminidase from B. alvei using mannan-Sepharose affinity chromatography. By using this method, endo-beta-N-acetylglucosaminidase was purified 3300-fold with 85% yield from the crude enzyme obtained by ammonium sulfate precipitation of the culture medium. The molecular weight of this enzyme was estimated to be about 66 000 by gel filtration. When using (Man)6(GlcNAc)2-Asn-Dns as substrate, the optimal activity occurs at pH 6.5 with Km of 1.9 mM. The action of endo-beta-N-acetylglucosaminidase toward several glycopeptides was also studied.

Acetylglucosaminidase↗
PubMed1983

Occurrence of a new hematoside in the kidney of guinea pig.

We have isolated a new hematoside from guinea pig kidney. Like the usual hematoside (II3NeuAc LacCer), isolated from human erythrocytes, this new hematoside contained glucose, galactose, and N-acetylneuraminic acid in an equimolar proportion. By thin-layer chromatography (TLC), however, it migrated faster than the usual hematoside. After mild alkaline hydrolysis the TLC mobility of this ganglioside became identical to that of the usual hematoside. The sialic acid in this ganglioside was susceptible to Clostridial neuraminidase. Based on TLC mobility and the results of periodate oxidation, the sialic acid of the new hematoside was identified as 9-O-acetyl-N-acetylneuraminic acid. Therefore, the structure of this new hematoside is 9-O-Ac-NeuAc alpha 2 leads to 3Gal beta 1 leads to 4GLc beta 1 leads to 1'Cer.

Animals↗
PubMed1983

The protein activator specific for the enzymic hydrolysis of GM2 ganglioside in normal human brain and brains of three types of GM2 gangliosidosis.

In order to understand the etiology of Type AB GM2 gangliosidosis, we have purified and characterized the activator protein (GM2 activator) specific for the enzymic hydrolysis of GM ganglioside from normal human brain. The purified activator from human brain moved as one major protein band in various electrophoretic systems. We have also prepared the antiserum against this activator. The levels and the nature of GM2 activator and beta-hexosaminidase A were examined in the brains of five cases of GM2 gangliosidosis-one Type B, two O, and two Type AB. We found that the levels of GM2 activator in both Type B and Type O cases were markedly elevated, and that the two Type AB cases were the results of different causes. One case had a defective beta-hexosaminidase A and an elevated level of GM2 activator. Although this defective beta-hexosaminidase A could hydrolyze synthetic substrates, it was inactive in the cleavage of natural glycosphingolipids in the presence of the GM2 activator. It could, however, hydrolyze asialo-GM2 and GbOse4Cer in the presence of sodium taurodeoxycholate. The other case had normal beta-hexosaminidase A, but had a very low level of GM2 activator when analyzed by in vitro assay, suggesting the deficiency of this activator. By immunoelectrophoresis, this case was found to be completely devoid of the protein that cross-reacts with the antiserum against the GM2 activator.

Brain↗
PubMed1983

Differentiation of two variants of type-AB GM2-gangliosidosis using chromogenic substrates.

Two variants of type-ABGM2-gangliosidosis can be distinguished by using p-nitrophenyl-6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside (PNP-GlcNAc-6-SO4) as substrate. One of the variants is caused by a deficiency of the activator for the hydrolysis of GM2-ganglioside. The beta-hexosaminidase A from this variant has a normal activity toward both PNP-GlcNAc and PNP-GlcNAc-6-SO4. A second variant caused by a defect in the enzyme, beta-hexosaminidase A, exhibits severely attenuated activity toward PNP-GlcNAc-6-SO4 but normal activity toward PNP-GlcNAc.

Acetylglucosamine↗
PubMed1983

Presence of activator proteins for the enzymic hydrolysis of GM1 and GM2 gangliosides in normal human urine.

Normal human urine has been found to contain activator proteins that stimulate the enzymic hydrolysis of GM1 and GM2 gangliosides. These two activators were partially purified by Sephadex G-200 filtration and DEAE-Sephadex A-50 chromatography. The presence of these two activators was assayed by demonstrating the stimulation of the in vitro hydrolysis of GM1 and GM2 gangliosides. As little as 50 ml of urine is sufficient to detect the presence of these two activators. The crude activator preparation from normal urine was also found to stimulate the hydrolysis of galactosylceramide sulfate by arylsulfatase A.

Adult↗
PubMed1982

Carbohydrate structure of human fibrinogen. Use of 300-MHz 1H-NMR to characterize glycosidase-treated glycopeptides.

The carbohydrate composition of fibrinogen and constituent S-carboxymethylated chains was determined. All the carbohydrate on the parent molecule could be accounted for on the B beta and gamma chains. The A alpha chain was found to be devoid of carbohydrate. Glycopeptides were prepared from fibrinogen, B beta, and gamma chains by pronase digestion and subsequent chromatography on Sephadex G-50. The 300-MHz 1H-NMR spectra of glycopeptides from all three sources were consistent with biantennary type oligosaccharide chains. The glycopeptides resulting from exoglycosidase digestions were examined with 300-MHz 1H-NMR spectroscopy, and the changes in the signals caused by selective removal of sugar residues were studied. This technique enabled us to assign each anomeric proton to the corresponding monosaccharide unit independent of previous work. Our results indicate that fibrinogen glycopeptides are of the biantennary type, and are in complete agreement with the previously reported peak assignments (Vliegenthart, J. F. G., van Halbeek, H., and Dorland, L. (1981) Pure Appl. Chemn. 53, 45-77). Affinity chromatography of the glycopeptides on concanavalin A-Sepharose also showed that the glycopeptides from fibrinogen are greater than 95% biantennary oligosaccharide chains. Based on carbohydrate composition, 1H-NMR spectroscopy, sequential exoglycosidase digestion, and gas chromatography-mass spectrometry of derived partially methylated alditol acetates, we propose that fibrinogen contains four oligosaccharide chains of the structure shown below. (formula, see text)

Carbohydrate Sequence↗
PubMed1982

Lewis blood group fucolipids and their isomers from human and canine intestine.

Glycolipids containing fucose linked to N-acetylglucosamine were isolated and characterized from 14 individual human and 13 individual dog intestines. From 8 of the dog intestines, Lewis a isomer fucolipids were isolated, all identical and having the structure Gal(beta 1 leads to 4)[Fuc alpha 1 leads to 3]GlcNAc(beta 1 leads to 3)Gal(beta 1 leads to 4)Glc-ceramide. Lewis b isomer fucolipids were isolated from 12 of the intestines, all identical and having the structure Fuc(alpha 1 leads to 2)Gal(beta 1 leads to 4)[Fuc alpha 1 leads to 3]GlcNAc(beta 1 leads to 3)Gal(beta 1 leads to 4)Glc-ceramide. Lewis a-active glycolipids were isolated as the sole major fucolipid in 6 of the human intestines and differed from the canine isomer only in the position of the linkage of galactose to N-acetylglucosamine, having the beta 1 leads to 3 (type 1) rather than the beta 1 leads to 4 (type 2) linkage. Lewis b-active fucolipids were isolated from 8 human intestines and differed from their canine isomer only in that they, too, had the type 1 rather than the type 2 oligosaccharide chain. Lewis a and b glycolipid isomers commonly co-existed in canine intestine as major fucolipids whereas Lewis a and b glycolipids did not so co-exist in human intestine. In all of the fucolipids, only hydroxylated fatty acids were present and phytosphingosine and sphingosine were the predominant long chain bases. These findings are of interest in the biosynthesis of these substances and in their genetic expression.

Animals↗