Isolation and characterization of the major .beta.-N-acetyl-D-glucosaminidase from human plasma

Verpoorte, Jacob A.

doi:10.1021/bi00701a023

Cited by 38 publications

(10 citation statements)

References 44 publications

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“…The glycoprotein nature of the NAG from several human tissues and serum has been established [15,16], Isoenzyme A contains sialic acid [15], Studies on human fibroblast (p < 0.0025). In the electrophoretic analysis both band A (more acidic, faster moving) and band B were visible in each sample.…”

Section: Discussionmentioning

confidence: 99%

Serum β-TV-Acetylglucosaminidase Isoenzymes in Insulin-Dependent Diabetes mellitus

Pitkänen¹,

Järvisalo²,

Harjanne³

et al. 1982

Enzyme

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Section: Discussionmentioning

confidence: 99%

Serum β-TV-Acetylglucosaminidase Isoenzymes in Insulin-Dependent Diabetes mellitus

Pitkänen¹,

Järvisalo²,

Harjanne³

et al. 1982

Enzyme

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“…Reference glycolipids were obtained by acid hydrolysis of mixed brain gangliosides [13], and characterized by T L C followed by gas-liquid chromatography [41]. p-D-N-Acetylhexosaminidase A was partially purified [44] from the patient's and normal plasma, by salt fractionation and DEAE-Sephadex chromatography [50]. p-D-N-Acetylhexosaminidase activity was measured [22] with 4-methylumbellileryl (4-MU) derivatives of p-D-N-acetylglucosaminide (GlcNAc) or p-D-N-acetylgalactosaminide (GalNAc) [51], and protein was determined by the method of Lowry et al [15].…”

Section: Imatel-ials and Methodsmentioning

confidence: 99%

A New Variant of Sandhoff's Disease

et al. 1974

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ExtractA 28-month-old Negro male with atypical Sandhoff's disease (GM2 gangliosidosis, type 2) is described. Clinical presentation closely resembled Sandhoff's disease. The appendiceal neuron cytoplasm was distended with periodic acid-Schiff (PAS)-positive material. Hepatic glycolipid N-acetylneuraminic acid was elevated 1.5-fold, chiefly because of increased GM2. Neutral glycolipid hexose was elevated twofold, mainly because of a component with globoside-like chromatographic properties. Unlike patients with Sandhops disease, the child had total 0-D-N-acetylhexosaminidase activity 20-24% of normal in serum and plasma and 7-1 1% of normal in leukocytes and cultured fibroblasts, but activity in liver (<2y0 of normal) was similar to that in Sandhoff's disease. In all tissues examined, >95% of the activity was heat denaturable, corresponding in electrophoretic mobility to the heat-denaturable component of normal tissues. The decrease in heat-denaturable activity with time and the pH optimum were similar in patient and control crude tissue preparations and partially purified preparations of plasma hexosaminidase A. In the child's mother, total hexosaminidase activity was approximately 50y0 of mean control values in serum, plasma, and leukocytes, and serum hexosaminidase A was relatively increased. This finding, together with the death of at least three other children in the kindred from an apparently phenotypically similar illness, indicates the inherited nature of the disorder. SpeculationAppreciable hexosaminidase activity in serum and plasma and its marked deficiency in liver suggest that the active form (or forms) in serum and plasma differ from those in solid tissues, and that the mutation in the case described has altered the hexosaminidases in a manner that virtually abolishes their activity in solid tissues but permits their partial activity in serum and plasma.

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“…Without a stabilizing agent, 5 µg•mL −1 PhNah20A was completely inactivated within 5 min at 50 • C, while 50% and 3% activity were retained after 20 min and 4 h, respectively, in 0.5% BSA ( Figure S5), and activity was fully retained after 4 d at 37 • C. β-NAHAs from E. coli [57], Prunus serotina [58], Bos taurus [59], Hordeum vulgare [60] and Streptomyces plicatus [61] were similarly found to lose activity by dilution. BSA has been identified as an activating compound to some β-NAHAs, e.g., from Mus musculus [41] and human plasma [42].…”

Section: Enzyme Stabilitymentioning

confidence: 99%

“…Murine cytosolic β-NAHA shows K M = 0.25 mM on pNPGalNAc, which it preferred over pNPGlcNAc [41]. Similarly, human plasma and pig brain β-NAHAs have a lower K M for pNPGalNAc of 0.17 mM and 0.2 mM, respectively [42,43]. Interestingly, salt-tolerant HJ5Nag from Microbacterium sp.…”

Section: Introductionmentioning

confidence: 99%

Identification and Characterization of a β-N-Acetylhexosaminidase with a Biosynthetic Activity from the Marine Bacterium Paraglaciecola hydrolytica S66T

Visnapuu

Tezé

Kjeldsen

et al. 2020

IJMS

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β-N-Acetylhexosaminidases are glycoside hydrolases (GHs) acting on N-acetylated carbohydrates and glycoproteins with the release of N-acetylhexosamines. Members of the family GH20 have been reported to catalyze the transfer of N-acetylglucosamine (GlcNAc) to an acceptor, i.e., the reverse of hydrolysis, thus representing an alternative to chemical oligosaccharide synthesis. Two putative GH20 β-N-acetylhexosaminidases, PhNah20A and PhNah20B, encoded by the marine bacterium Paraglaciecola hydrolytica S66T, are distantly related to previously characterized enzymes. Remarkably, PhNah20A was located by phylogenetic analysis outside clusters of other studied β-N-acetylhexosaminidases, in a unique position between bacterial and eukaryotic enzymes. We successfully produced recombinant PhNah20A showing optimum activity at pH 6.0 and 50 °C, hydrolysis of GlcNAc β-1,4 and β-1,3 linkages in chitobiose (GlcNAc)2 and GlcNAc-1,3-β-Gal-1,4-β-Glc (LNT2), a human milk oligosaccharide core structure. The kinetic parameters of PhNah20A for p-nitrophenyl-GlcNAc and p-nitrophenyl-GalNAc were highly similar: kcat/KM being 341 and 344 mM−1·s−1, respectively. PhNah20A was unstable in dilute solution, but retained full activity in the presence of 0.5% bovine serum albumin (BSA). PhNah20A catalyzed the formation of LNT2, the non-reducing trisaccharide β-Gal-1,4-β-Glc-1,1-β-GlcNAc, and in low amounts the β-1,2- or β-1,3-linked trisaccharide β-Gal-1,4(β-GlcNAc)-1,x-Glc by a transglycosylation of lactose using 2-methyl-(1,2-dideoxy-α-d-glucopyrano)-oxazoline (NAG-oxazoline) as the donor. PhNah20A is the first characterized member of a distinct subgroup within GH20 β-N-acetylhexosaminidases.

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Isolation and characterization of the major .beta.-N-acetyl-D-glucosaminidase from human plasma

Cited by 38 publications

References 44 publications

Serum β-TV-Acetylglucosaminidase Isoenzymes in Insulin-Dependent Diabetes mellitus

Serum β-TV-Acetylglucosaminidase Isoenzymes in Insulin-Dependent Diabetes mellitus

A New Variant of Sandhoff's Disease

Identification and Characterization of a β-N-Acetylhexosaminidase with a Biosynthetic Activity from the Marine Bacterium Paraglaciecola hydrolytica S66T

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