Biochemical and Immunological Studies on Two Distinct Ganglioside-Hydrolyzing Sialidases from the Particulate Fraction of Rat Brain1

Miyagi, Taeko; Sagawa, Junji; Konno, K.; Handa, Shizuo; Tsuiki, Shigeru

doi:10.1093/oxfordjournals.jbchem.a123126

Cited by 83 publications

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“…Sialidases or neuraminidases (EC 3.2.1.18) are hydrolytic enzymes that remove sialic acid residues from gangliosides and other natural substrates. Among these proteins, plasma membrane-associated sialidase has been described in different tissues (15)(16)(17) and cell types (18 -20), and a form linked to the membrane via glycosylphosphatidylinositol anchor has also been identified (21,22). Evidence of a possible involvement of this enzyme in the regulation of the sialic acid levels of the cell surface has been produced.…”

Section: Glycosphingolipids (Gsls)mentioning

confidence: 99%

The Plasma Membrane-associated Sialidase MmNEU3 Modifies the Ganglioside Pattern of Adjacent Cells Supporting Its Involvement in Cell-to-Cell Interactions

Papini

Anastasia

Tringali

et al. 2004

Journal of Biological Chemistry

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131

142

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We describe herein the enzyme behavior of MmNEU3, the plasma membrane-associated sialidase from mouse (Mus musculus). MmNEU3 is localized at the plasma membrane as demonstrated directly by confocal microscopy analysis. In addition, administration of the radiolabeled ganglioside GD1a to MmNEU3-transfected cells, under conditions that prevent lysosomal activity, led to its hydrolysis into ganglioside GM1, further indicating the plasma membrane topology of MmNEU3. Metabolic labeling with [1-3 H]sphingosine allowed the characterization of the ganglioside patterns of COS-7 cells. MmNEU3 expression in COS-7 cells led to an extensive modification of the cell ganglioside pattern, i.e. GM3 and GD1a content was decreased to about one-third compared with mock-transfected cells. At the same time, a 35% increase in ganglioside GM1 content was observed. Mixed culture of MmNEU3-transfected cells with [1-3 H]sphingosine-labeled cells demonstrates that the enzyme present at the cell surface is able to recognize gangliosides exposed on the membrane of nearby cells. Under these experimental conditions, the extent of ganglioside pattern changes was a function of MmNEU3 transient expression. Overall, the variations in GM3, GD1a, and GM1 content were very similar to those observed in the case of [1-3 H]sphingosine-labeled MmNEU3-transfected cells, indicating that the enzyme mainly exerted its activity toward ganglioside substrates present at the surface of neighboring cells. These results indicate that the plasma membrane-associated sialidase MmNEU3 is able to hydrolyze ganglioside substrates in intact living cells at a neutral pH, mainly through cell-to-cell interactions. Glycosphingolipids (GSLs)1 expressed at the cell surface are well known as modulators of several aspects of signal transduction processes involved in the control of cell proliferation, survival, and differentiation (1). GSLs in the plasma membrane are able to interact laterally with other membrane molecules modulating their properties (cis-interactions). Lipid rafts or membrane microdomains result from dynamic clustering of sphingolipids and cholesterol to form the so-called sphingolipid-enriched domain (SED) or lipid rafts (2). These structures move within the fluid bilayer and function as platforms for the attachment of proteins when membranes are moved around the cell and during signal transduction (3, 4). In addition, the expression pattern of GSLs in several cells and tissues undergoes deep changes during development and neoplastic transformation (5). These events are usually characterized by dramatic changes in cell recognition, suggesting that GSLs as cell surface antigens could play relevant roles as receptor sites in cell-cell recognition (trans-interaction). The receptor role of GSLs has been hypothesized in the case of microbial infections based on their ability to interact with bacterial toxins and microbial lectins (6, 7). Conformational analysis confirms that the orientation of the oligosaccharide chains of glycolipids at the cell surface complies wit...

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Section: Glycosphingolipids (Gsls)mentioning

confidence: 99%

The Plasma Membrane-associated Sialidase MmNEU3 Modifies the Ganglioside Pattern of Adjacent Cells Supporting Its Involvement in Cell-to-Cell Interactions

Papini

Anastasia

Tringali

et al. 2004

Journal of Biological Chemistry

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131

142

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“…These developmental differences, together with property differences (for example the optimal pH) may be in favour of the concept [22,23] that the various…”

Section: Ultrastructural Analysismentioning

confidence: 99%

Occurrence of sialidase activity in two distinct and highly homogeneous populations of lysosomes prepared from the brain of developing mouse

Fiorilli¹,

Siniscalco²,

Chiarini³

et al. 1991

FEBS Letters

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Lillht and l~eavy lyxo=~ome~ of mou~,= forebrain were s~=parated from ¢a¢11 other hy ¢enlrtfugalion ~n a Pereoll gradient, Light lysosomes were then freed from mitoehondria and membrane~ by stterose density Bl'adieat eentriful~atica and further puritied by floalatton-eentrifuBalion on a sucrose tlradient, "['he final prcparation~ of liiht and heavy ly~osom~, Fairly homogenous, carried sialidase activity, assayed on MU-NeuA¢. The optimal NI wa~ 4,0 and 4.2, the apparent g.~ value 2.8 x 10 =~ M and 4,2 x i0 "~ M and tire apparent 1~ value 0.11 and 0.47 mU, mj =~ protoin, for the liBht and heavy ly~to~ome sialidas¢, respectively, From 4 day~ t0 adulthood the sp,.~ific ac~ivily of the li~,ht and heavy iysosome lialidase increased 3.fold and I

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“…Based on their substrate specificities, immunological properties and subcellular localization, mammalian sialidases have been classified into four types by Miyagi et al [8][9][10] : cytosolic [8], intralysosomal [9], lysosomal-membrane-associated [10] and plasma-membraneassociated [10] sialidases. The first molecular cloning of a mammalian sialidase was reported by Miyagi et al [11] in 1993 for rat skeletal-muscle cytosolic-type sialidase.…”

Section: Introductionmentioning

confidence: 99%

Degradation of GM1 and GM2 by mammalian sialidases

Moriya

et al. 2001

Biochemical Journal

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In mammalian tissues, the pathway known for the catabolism ofGM1[Galβ3GalNAcβ4(Neu5Acα3)Galβ4GlcCer;where Cer is ceramide] is the conversion of this ganglioside into GM2 [GalNAcβ4(Neu5Acα3)Galβ4GlcβCer] by β-galactosidase followed by the conversion of GM2 into GM3 (Neu5Acα3Galβ4GlcβCer) by β-N-acetylhexosaminidase A (Hex A). However, the question of whether or not GM1 and GM2 can also be respectively converted into asialo-GM1 (Galβ3GalNAcβ4Galβ4GlcCer; GA1) and asialo-GM2 (GalNAcβ4Galβ4GlcβCer, GA2) by mammalian sialidases has not been resolved. This is due to the fact that sialidases purified from mammalian tissues always contained detergents that interfered with the in vitro hydrolysis of GM1 and GM2 in the presence of an activator protein. The mouse model of human type B Tay–Sachs disease created by the disruption of the Hexa gene showed no neurological abnormalities, with milder clinical symptoms than the human counterpart, and the accumulation of GM2 in the brains of affected mice was only limited to certain regions [Sango, Yamanaka, Hoffmann, Okuda, Grinberg, Westphal, McDonald, Crawley, Sandhoff, Suzuki and Proia (1995) Nat. Genet. 11, 170–176]. These results suggest the possible presence of an alternative catabolic pathway (the GA2 pathway) in mouse to convert GM2 into GA2 by sialidase. To show the existence of this pathway, we have used recombinant mammalian cytosolic sialidase and membrane-associated sialidase to study the desialylation of GM1 and GM2. We found that the mouse membrane-bound sialidase was able to convert GM1 and GM2 into their respective asialo-derivatives in the presence of human or mouse GM2 activator protein. The cytosolic sialidase did not exhibit this activity. Our results suggest that, in vivo, the stable NeuAc of GM1 and GM2 may be removed by the mammalian membrane-associated sialidase in the presence of GM2 activator protein. They also support the presence of the GA2 pathway for the catabolism of GM2 in mouse.

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Biochemical and Immunological Studies on Two Distinct Ganglioside-Hydrolyzing Sialidases from the Particulate Fraction of Rat Brain1

Cited by 83 publications

References 0 publications

The Plasma Membrane-associated Sialidase MmNEU3 Modifies the Ganglioside Pattern of Adjacent Cells Supporting Its Involvement in Cell-to-Cell Interactions

The Plasma Membrane-associated Sialidase MmNEU3 Modifies the Ganglioside Pattern of Adjacent Cells Supporting Its Involvement in Cell-to-Cell Interactions

Occurrence of sialidase activity in two distinct and highly homogeneous populations of lysosomes prepared from the brain of developing mouse

Degradation of GM1 and GM2 by mammalian sialidases

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