The demonstration of multiple heat stable forms of N-acetyl-β-glucosaminidase in normal human serum

Price, Robert G.; Dance, N.

doi:10.1016/0005-2795(72)90142-0

Cited by 148 publications

(42 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Except for the marked increase in activity, these bands appeared to be identical with the faster moving bands (A and I 2 [25]) routinely seen in normal serum samples (Fig. 1) [10]. In these samples, however, the I 2 band, not the A band, accounted for the major proportion of the activity.…”

Section: Resultssupporting

confidence: 54%

See 1 more Smart Citation

Mucolipidosis III (Pseudo-Hurler Polydystrophy): Multiple Lysosomal Enzyme Abnormalities in Serum and Cultured Fibroblast Cells

et al. 1973

View full text Add to dashboard Cite

Section: Resultssupporting

confidence: 54%

“…This belief is based on the observation that the I 2 activity is stable under the conditions employed for the heat denaturation determination while the A activity is destroyed [10].…”

Section: Discussionmentioning

confidence: 99%

Mucolipidosis III (Pseudo-Hurler Polydystrophy): Multiple Lysosomal Enzyme Abnormalities in Serum and Cultured Fibroblast Cells

et al. 1973

View full text Add to dashboard Cite

“…21 -231 but the active sites for these respective two substrates might be distinct [24, [8,[27][28][29] seem to be identical to form P characterized in the serum of pregnant women [30 - [34,351 or alternatively identical to Hex B [36, 371. Hex S [38,39], a minor acidic form (PI about 4.5) present in normal liver [35] and in Sandhoff disease tissues [36,401 hydrolyses substrate containing both the P-GlcNAc and @-GalNAc linkages.…”

mentioning

confidence: 99%

Molecular Forms of β‐N‐Acetylhexosaminidase in Epstein‐Barr Virus‐Transformed Lymphoid Cell Lines from Normal Subjects and Patients with Tay‐Sachs Disease

Salvayre

Maret

Nègre

et al. 1983

European Journal of Biochemistry

View full text Add to dashboard Cite

In whole leukocytes and in lymphocytes from normal subjects, the percentage activity of heat‐stable β‐N‐acetylhexosaminidase (30 ± 5% and 45 ± 5%, respectively) was higher than in the transformed lymphoid cell line (19 ± 3%). In Tay‐Sachs transformed cells as well as non‐transformed lymphocytes, β‐N‐acetylhexosaminidase was almost completely heat‐stable (95–98%). In the transformed cells from normal subjects, the β‐N‐acetylhexosaminidase B (Hex B) activity (5% of total) was significantly lower than in blood lymphocytes (average 25–30% of total activity), whereas Hex A and Hex I were similar in the either cell type. Blood lymphocytes and lymphoid cell lines established from a Tay‐Sachs patient lacked heat‐labile Hex A and expressed high heat‐stable Hex I and Hex B activities (3–6‐fold). After neuraminidase treatment, Hex A peak sharpened while Hex I peaks switched to higher pI than normal Hex I, in the region of Hex B. PreHex A/S pI was not affected. Hydrolytic properties using the both substrates (4‐methylumbelliferyl‐2‐acetamido‐2‐deoxy‐β‐d‐glucopyranoside and 4‐methylumbelliferyl‐2‐acetamido‐2‐deoxy‐β‐d‐galactopyranoside) of each molecular form were similar in transformed and non‐transformed cells. Data derived from the use of a mixture of substrates were consistent with the model which proposes a common active site for either substrate in the case of preHex A, Hex B and Hex I, but not for Hex A.Thus Epstein‐Barr virus‐transformed lymphoid cell lines represent an accurate model system for studies on Tay‐Sachs disease.

show abstract

“…Several of these mammalian tissues also contained at least two or three other b-N-acetylglucosaminidase activities which eluted from DEAE-cellulose columns like the octopus PII, PITI and p l V (Table 1) [24,27,28]. While in the octopus PI represented about 80% of the total activity, in mammalian tissues p-N-acetylglucosaminidase A, possibly corresponding to [W, was the most prominent species [I, 8, 24,25,27-291.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a β‐N‐Acetylglucosaminidase from Octopus vulgaris

Ceccarini

D’Aniello

Cacace³

et al. 1983

European Journal of Biochemistry

View full text Add to dashboard Cite

We have purified a p-N-acetylglucosaniinidase from the hepatopancreas of the octopus which we have called PI. The enzyme was homogeneous as judged by Sephadex column chromatography, isoelectric focusing, non-denaturing gel electrophoresis at two different pH and with sodium dodecyl sulphate/polyacrylamide gel electrophoresis. The native protcin has an apparent molecular weight of 120000 and we can conclude that it is a tetramer made up of two CI and two P subunits with apparent M , of 27000 and 34000, respectively. Using N M R spectroscopy we have examined the specificity of fi1 and have established that the enzyme hydrolyses the p1,4 linkage of N-acetylglucosamine but at only a specific site of the substrates used, two glycopeptides isolated from ovalbumin. To our knowledge this is the first known exoglycosidase which has both linkage and site specificity. P-N-Acetylglucosaminidase (EC 3.2.1.30) has been studied and purified from a variety of sources [1 -81. The enzyme is an exoglycosidase capable of removing N-acetylglucosamine from oligosaccharides and glycopeptides or glycoproteins. All of the hexosaminidases so far studied do not appear to recognize specific glycone substrates of known structure; however, they require the p configuration of the sugar. In a survey for specific glycosidases initiated by one of us (C.C.), the hepatopancreas of octopus was screened for glycosidasc activities [9]. This organ was found to be an extremely rich source of hexosaminidase activity and in this paper we report the purification and the biochemical properties of a p-Nacetylglucosaminidase. We also demonstrate that this enzyme is specific for the Pl,4 linkage of N-acetylglucosamine situated at a unique site of ovalbumin glycopeptide A [lo, 111. We have established this by high-resolution 'H-NMR spectroscopy. To our knowledge this exquisite specificity has never been previously reported for an exoglycosidase and suggests that glycosidase activity can be as finely regulated as is the biosynthesis of oligosaccharide fine structure.

show abstract

The demonstration of multiple heat stable forms of N-acetyl-β-glucosaminidase in normal human serum

Cited by 148 publications

References 14 publications

Mucolipidosis III (Pseudo-Hurler Polydystrophy): Multiple Lysosomal Enzyme Abnormalities in Serum and Cultured Fibroblast Cells

Mucolipidosis III (Pseudo-Hurler Polydystrophy): Multiple Lysosomal Enzyme Abnormalities in Serum and Cultured Fibroblast Cells

Molecular Forms of β‐N‐Acetylhexosaminidase in Epstein‐Barr Virus‐Transformed Lymphoid Cell Lines from Normal Subjects and Patients with Tay‐Sachs Disease

Purification and Characterization of a β‐N‐Acetylglucosaminidase from Octopus vulgaris

Contact Info

Product

Resources

About