Isolation of β-N-Acetylhexosaminidase, β-N-Acetylglucosaminidase, and β-N-Acetylgalactosaminidase from Calf Brain<sup>*</sup>

Frohwein, Yaacov Zvi; Gátt, Shimon

doi:10.1021/bi00861a018

Cited by 112 publications

(34 citation statements)

References 25 publications

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“…Similar to previous purifications in liver [10], spleen [8], kidney [3], and brain [4], centrifugation, dialysis, ion exchange and gel filtration chromatog raphy resulted in increases of hexosaminidase activities. In preliminary experiments, crude lens homogenates were precipitated with NH4 S 04 in concentrations ranging from 25 to 75% saturation, but only 2-to 4-fold increases in activities were found after dialysis.…”

Section: Discussionsupporting

confidence: 79%

“…Hexosaminidases, (V-acetyl-^-D-glucosaminidase and 7V-acetyl-/?-D-galactosaminidase, have been purified from several mammalian tissues, includ ing liver [10], spleen [8], kidney [3] and brain [4], W eissmann et al have [10] shown that beef liver glucosaminidase liberates ¿V-acetylglucosamine from the terminal position of oligosaccharides while F r o h w ein and G att [4] have demonstrated similar hydrolytic functions for glucosaminidase and galactosaminidase on oligosaccharides and sphingolipids of calf brain. Separation of splenic glucosaminidase into heat-labile (hexosaminidase A) and heat-stable (hexosaminidase B) forms was accomplished by R obinson and Stir l in g [8], and D ance et al [3] demonstrated two hexosaminidase forms in human kidney.…”

Section: Introductionmentioning

confidence: 99%

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Lens Hexosaminîdases: Purification and Properties

Cotlier¹,

Carlin²

1976

Ophthalmic Res

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Lens hexosaminidase(s), N-acetyl-β-D-glucosaminidase and N-acetyl-β-D-galactosaminidase, were purified from rabbit lens capsule-epithelium by centrifugation, dialysis and column chromatography on DEAE-cellulose ans Sephadex G-200. pH distribution curves and kinetic constants (K_m and V_max) were determined in the fractions purified approximately 700-fold. Lens glucosaminidase and galactosaminidase could not be separated from each other, thus indicating that one enzyme was present but unable to distinguish the C₄ position of aminosugars.

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Section: Discussionsupporting

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Lens Hexosaminîdases: Purification and Properties

Cotlier¹,

Carlin²

1976

Ophthalmic Res

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“…Restriction enzymes, T4 DNA ligase, globotetraosylceramide (Gb4Cer), 3 and globotriaosylceramide (Gb3Cer) were from Wako Pure Chemical Inc. Ltd. The crude ganglioside mixture was prepared from bovine brain as described previously (13), and GM2, asialo-GM1 (GA1), asialo-GM2 (GA2; gangliotriaosylceramide), lactosylceramide (LacCer), and glucosylceramide (GlcCer) were prepared from the crude ganglioside mixture using GSL-degrading enzymes from Paenibacillus sp.…”

Section: Methodsmentioning

confidence: 99%

“…Among the ␤-HEXs, an enzyme that hydrolyzes the nonreducing terminal ␤-GalNAc residue much faster than the ␤-GlcNAc residue is named ␤-N-acetylgalactosaminidase (␤-NGA; EC 3.2.1.53). ␤-NGA activities have been detected in bovine brain (3), rat brain (4), and bacteria (5); however, ␤-NGA has not yet been fully characterized. Recently, a nucleocytoplasmic neutral ␤-HEX (named ␤-HEX D) was cloned from humans and mice (6) as a homolog of Caenorhabditis elegans ␤-HEX (7).…”

mentioning

confidence: 99%

Molecular Cloning and Catalytic Mechanism of a Novel Glycosphingolipid-degrading β-N-Acetylgalactosaminidase from Paenibacillus sp. TS12

Sumida

Fujimoto

Ito

2011

Journal of Biological Chemistry

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We report here the molecular cloning, characterization, and catalytic mechanism of a novel glycosphingolipid-degrading ␤-N-acetylgalactosaminidase (␤-NGA) from Paenibacillus sp. TS12 (NgaP). Consisting of 1034 putative amino acid residues, NgaP shares no sequence similarity with known proteins. Recombinant NgaP, expressed in Escherichia coli, cleaved the nonreducing terminal ␤-GalNAc residues of gangliotriaosylceramide and globotetraosylceramide. The enzyme hydrolyzed para-nitrophenyl-␤-N-acetylgalactosaminide ϳ100 times faster than para-nitrophenyl-␤-N-acetylglucosaminide. GalNAc thiazoline, an analog of the oxazolinium intermediate and potent inhibitor for enzymes adopting substrate-assisted catalysis, competitively inhibited the enzyme. The K i of the enzyme for GalNAc thiazoline was 1.3 nM, whereas that for GlcNAc thiazoline was 46.8 M. Comparison of the secondary structure with those of known enzymes exhibiting substrate-assisted catalysis and point mutation analysis indicated that NgaP adopts substrate-assisted catalysis in which Glu-608 and Asp-607 could function as a proton donor and a stabilizer of the 2-acetamide group of the ␤-GalNAc at the active site, respectively. These results clearly indicate that NgaP is a ␤-NGA showing substrateassisted catalysis. This is the first report describing the molecular cloning of a ␤-NGA adopting substrate-assisted catalysis.

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“…1,2-Bisepi-valienamine 153 acts as a -mannosidase [321] inhibitor whereas 2-acetamido-2-deoxy-1-epi-valienamine 154 has been shown [322] to inhibit various -hexosaminidases, enzymes involved in the hydrolysis of terminal N-acetyl-D-hexosamine residues in N-acetyl--D-hexosaminides [323][324][325].…”

Section: International Journal Of Carbohydrate Chemistrymentioning

confidence: 99%

Biosynthesis and Biological Activity of Carbasugars

Roscales

Plumet

2016

International Journal of Carbohydrate Chemistry

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The first synthesis of carbasugars, compounds in which the ring oxygen of a monosaccharide had been replaced by a methylene moiety, was described in 1966 by Professor G. E. McCasland's group. Seven years later, the first true natural carbasugar (5a-carba-R-D-galactopyranose) was isolated from a fermentation broth of Streptomyces sp. MA-4145. In the following decades, the chemistry and biology of carbasugars have been extensively studied. Most of these compounds show interesting biological properties, especially enzymatic inhibitory activities, and, in consequence, an important number of analogues have also been prepared in the search for improved biological activities. The aim of this review is to give coverage on the progress made in two important aspects of these compounds: the elucidation of their biosynthesis and the consideration of their biological properties, including the extensively studied carbapyranoses as well as the much less studied carbafuranoses.

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Isolation of β-N-Acetylhexosaminidase, β-N-Acetylglucosaminidase, and β-N-Acetylgalactosaminidase from Calf Brain^*

Cited by 112 publications

References 25 publications

Lens Hexosaminîdases: Purification and Properties

Lens Hexosaminîdases: Purification and Properties

Molecular Cloning and Catalytic Mechanism of a Novel Glycosphingolipid-degrading β-N-Acetylgalactosaminidase from Paenibacillus sp. TS12

Biosynthesis and Biological Activity of Carbasugars

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Isolation of β-N-Acetylhexosaminidase, β-N-Acetylglucosaminidase, and β-N-Acetylgalactosaminidase from Calf Brain*

Cited by 112 publications

References 25 publications

Lens Hexosaminîdases: Purification and Properties

Lens Hexosaminîdases: Purification and Properties

Molecular Cloning and Catalytic Mechanism of a Novel Glycosphingolipid-degrading β-N-Acetylgalactosaminidase from Paenibacillus sp. TS12

Biosynthesis and Biological Activity of Carbasugars

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Isolation of β-N-Acetylhexosaminidase, β-N-Acetylglucosaminidase, and β-N-Acetylgalactosaminidase from Calf Brain^*