Interaction of Human Brain Neuraminidase with Tritium—Labelled Gangliosides

Veh, Rüdiger W.; Schauer, Roland

doi:10.1007/978-1-4615-9071-2_41

Cited by 17 publications

(4 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the brain, ganglioside-degrading sialidases have been described in particulate fractions and in myelin by a number of workers [12,[14][15][16][35][36][37][38][39]. Recently, a ganglioside sialidase with a substrate specificity different from ours was detected in nuclei of rat brain [40,411.…”

Section: Discussionmentioning

confidence: 85%

Partial Characterization and Enrichment of a Membrane‐Bound Sialidase Specific for Gangliosides from Human Brain Tissue

Kopitz

Sinz

Brossmer

et al. 1997

European Journal of Biochemistry

View full text Add to dashboard Cite

Gangliosides, constituents of surfaces of vertebrate cells, modulate important cellular functions. Ganglioside-specific sialidases that possibly control these processes have been observed in a number of tissues, but their characterization has proved difficult due to their low abundance and lability. Here we describe the partial isolation and characterization of a ganglioside sialidase from human brain grey matter. After membrane extraction with octylglucoside, the enzyme was purified about 1300-fold by ion-exchange, affinity and gel-permeation chromatographies. Although PAGE still showed several protein bands, specific photoaffinity labelling with iodinated S-N-acetyl-9-(4-azidosalicoylamido)-2,9-dideoxy-2,3-didehydroneuraminic acid identified a single polypeptide of 60 kDa likely to contain the active site of the sialidase. In the presence of 0.4 % octylglucoside, the purified sialidase desialylated gangliosides G,,, G,,,,, G,,, and G,,,, but was inactive towards G,,, GM2, colominic acid, sialyl-(a2-3)-lactose, 2-(4-methylumbelliferyl)-neuraminate, or the glycoprotein fetuin. The ganglioside sialidase activity was strongly inhibited by 2-deoxy-2,3-didehydro-N-acetylneuraminic acid, heparin and heparan sulfate. Because of its substrate and inhibitor profiles, the purified enzyme resembles the activity characterized previously in the plasma membrane of human neuroblastoma cells, but is distinct from a lysosomal activity. The purified brain sialidase thus appears to function in the selective desialylation of gangliosides with terminal sialic acid residues.

show abstract

Section: Discussionmentioning

confidence: 85%

Partial Characterization and Enrichment of a Membrane‐Bound Sialidase Specific for Gangliosides from Human Brain Tissue

Kopitz

Sinz

Brossmer

et al. 1997

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…The radiolabeled GM3 was used as a substrate for the sialidase assay. GD1a was radiolabeled in the sialic acid portion according to the method of Veh and Schauer (1978) with modifications. In brief, GD1a was oxidized by NaIO 4 .…”

Section: Sialidase Assaymentioning

confidence: 99%

Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells

Liour

Kapitonov

2000

J. Neurosci. Res.

View full text Add to dashboard Cite

Gangliosides are constituents of the cell membrane and are known to have important functions in neuronal differentiation. We employed an embryonal carcinoma stem cell line P19 as an in vitro model to investigate the expression of gangliosides during neuronal development. After treatment with retinoic acid, these cells differentiate synchronously into neuron‐like cells by a series of well‐defined events of development. We examined several aspects of ganglioside metabolism, including the changes of ganglioside pattern, the activities and gene expression of several enzymes at different stages of differentiation, and the distribution of gangliosides in differentiating neurons. Undifferentiated P19 cells express mainly GM3 and GD3. After P19 cells were committed to differentiation, the synthesis of complex gangliosides was elevated more than 20‐fold, coinciding with the stage of neurite outgrowth. During the maturation of differentiated cells, the expression of c‐series gangliosides was downregulated concomitantly with upregulation of the expression of a‐ and b‐series gangliosides. We also examined the distribution of gangliosides in differentiating neurons by confocal and transmission electron microscopy after cholera toxin B subunit and sialidase treatment. Confocal microscopic studies showed that gangliosides were distributed on the growth cones and exhibited a punctate localization on neurites and soma. Electron microscopic studies indicated that they also are enriched on the plasma membranes of neurites and the filopodia as well as on the lamellipodia of growth cones during the early stage of neurite outgrowth. Our data demonstrate that the expression of gangliosides in P19 cells during RA‐induced neuronal differentiation resembles that of the in vivo development of the vertebrate brain, and hence validates it as an in vitro model for investigating the function of gangliosides in neuronal development. J. Neurosci. Res. 62:363–373, 2000. © 2000 Wiley‐Liss, Inc.

show abstract

“…The K , values for NL and G,, changed significantly after the delipidation and extraction of the myelin enzyme. It was recently suggested that absorption of ganglioside substrates to membranes, which was used as the source of mammalian neuraminidase, might cause an elevation of the substrate concentrations in the microenvironment of the enzyme, resulting in an artificial decrease in the apparent K , values (Veh and Schauer, 1978). Since such an absorption probably occurs through hydrophobic interactions, it can be expected that the K , value with G,, would increase by the removal of myelin lipids from the myelin enzyme preparation.…”

Section: Discussionmentioning

confidence: 99%

“…Inhibition of enzyme activity by 2,3-dehydro-2-deoxy-N-acetylneuraminic acid 2,3-Dehydro-2-deoxy-N-acetylneuraminic acid is known to be a potent competitive inhibitor for neurarninidase (Veh and Schauer, 1978). The enzyme activities toward N L in myelin and microsomes were effectively inhibited by it (data not shown).…”

Section: Heat Stability Of the Myelin And Microsomal Enzymesmentioning

confidence: 93%

Further Characterization of a Myelin‐Associated Neuraminidase: Properties and Substrate Specificity

Saito

1986

Journal of Neurochemistry

View full text Add to dashboard Cite

A neuraminidase activity in myelin isolated from adult rat brains was examined. The enzyme activity in myelin was first compared with that in microsomes using N-acetylneuramin(a2-+3)lactitol (NL) as a substrate. In contrast to the microsomal neuraminidase which exhibited a sharp pH dependency for its activity, the myelin enzyme gave a very shallow pH activity curve over a range between 3.6 and 5.9. The myelin enzyme was more stable to heat denaturation (65°C) than the microsomal enzyme. Inhibition studies with a competitive inhibitor, 2,3-dehydro-2-deoxy-N-acetylneuraminic acid, showed the Ki value for the myelin neuraminidase to be about one-fifth of that for the microsomal enzyme (1.3 x M). The apparent K , values for the myelin and the microsomal enzyme were 1.3 x IO-'M and 4.3 x M, respectively. An enzyme preparation that was practically devoid of myelin lipids was then prepared and its substrate specificity examined. The "delipidated enzyme" could hydrolyze fetuin, NL, and ganglioside substrates, including G M l and GMz. When the

show abstract

Interaction of Human Brain Neuraminidase with Tritium—Labelled Gangliosides

Cited by 17 publications

References 19 publications

Partial Characterization and Enrichment of a Membrane‐Bound Sialidase Specific for Gangliosides from Human Brain Tissue

Partial Characterization and Enrichment of a Membrane‐Bound Sialidase Specific for Gangliosides from Human Brain Tissue

Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells

Further Characterization of a Myelin‐Associated Neuraminidase: Properties and Substrate Specificity

Contact Info

Product

Resources

About