Substrate Specificity and Inhibitor Studies of a Membrane-Bound Ganglioside Sialidase Isolated from Human Brain Tissue
-
C. Oehler
Abstract
A gangliosidespecific sialidase that controls cellular functions such as growth, differentiation, and adhesion has been observed in a variety of cells, but its characterization proved difficult due to firm membrane attachment and lability of the purified enzyme. Here we report on the specificity toward gangliosides and susceptibility to certain inhibitors of a ganglioside sialidase solubilized and purified 5100-fold from human brain. The sialidase removed terminal sialic acids from gangliosides GM3, GM4, GD3, GD2, GD1a, GD1b, GT1b and GQ1b, but was inactive toward gangliosides with sialic acid in a branching position (as in GM1 and GM2). LysoGM3 and GD1a were good substrates, too, whereas Oacetylation of the sialic acid as in 9-OacetylGD3 caused strongly reduced cleavage. The new influenza virus drug 4-guanidino-2-deoxy-2,3- dehydroNacetylneuraminic acid (Zanamivir) exhibited an IC50 value of about 710 5 M that was in the range of the classical sialidase inhibitor 2-deoxy-2,3-dehydro Nacetylneuraminic acid; the bacterial sialidase inhibitor 4-nitrophenyloxamic acid, however, was ineffective. The glycosaminoglycans heparan sulfate, heparin, chondroitin sulfates A and B, as well as dextran sulfate and suramin, were all strongly inhibitory, suggesting that glycosaminoglycans present on the cell surface or in the extracellular matrix may influence the ability of the sialidase to alter the ganglioside composition of the membrane.
Copyright © 2002 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Pigment Epithelium-Derived Factor (PEDF), a Serpin with Potent Anti-Angiogenic and Neurite Outgrowth-promoting Properties
- Mechanism of Inhibition of RNA Polymerase I Transcription by DNA-dependent Protein Kinase
- Evidence for Haploinsufficiency of the Human HNF1α Gene Revealed by Functional Characterization of MODY3-Associated Mutations
- In vitro and in vivo Stability of the 2ζ2 Protein Complex of the Broad Host-Range Streptococcus pyogenes pSM19035 Addiction System
- Effects of Antineoplastic Agents on Cytoplasmic and Membrane-Bound Heat Shock Protein 70 (Hsp70) Levels
- Characterization of the VPS10 Domain of SorLA/LR11 as Binding Site for the Neuropeptide HA
- Substrate Specificity and Inhibitor Studies of a Membrane-Bound Ganglioside Sialidase Isolated from Human Brain Tissue
- Specificity Determinants in the Interaction of Apolipoprotein(a) Kringles with Tetranectin and LDL
- Amylin-Induced Cytotoxicity Is Associated with Activation of Caspase-3 and MAP Kinases
- 2-Alkyl-3-Alkylamino-2H-Benzo- and Pyridothiadiazine 1,1-Dioxides: From K+ATP Channel Openers to Ca++ Channel Blockers?
- Pre-Loading of Chlorophyll Synthase with Tetraprenyl Diphosphate Is an Obligatory Step in Chlorophyll Biosynthesis
- Molecular and Immunological Characterization of Profilin from Mugwort Pollen
- The Archaeal Respiratory Supercomplex SoxM from S.acidocaldarius Combines Features of Quinole and Cytochrome c Oxidases
- Benzodiazepine Binding Studies on Living Cells: Application of Small Ligands for Fluorescence Correlation Spectroscopy
- Individual Recombinant Thyroglobulin Type-1 Domains Are Substrates for Lysosomal Cysteine Proteinases
- Characterization of Legumain
- Molecular Cloning and Chimerisation of an Inhibitory Anti-Cathepsin B Antibody and Its Expression in Chinese Hamster Ovary Cells
- Application of Peptides Containing the Cleavage Sequence of Pro-TNFα in Assessing TACE Activity of Whole Cells
Articles in the same Issue
- Pigment Epithelium-Derived Factor (PEDF), a Serpin with Potent Anti-Angiogenic and Neurite Outgrowth-promoting Properties
- Mechanism of Inhibition of RNA Polymerase I Transcription by DNA-dependent Protein Kinase
- Evidence for Haploinsufficiency of the Human HNF1α Gene Revealed by Functional Characterization of MODY3-Associated Mutations
- In vitro and in vivo Stability of the 2ζ2 Protein Complex of the Broad Host-Range Streptococcus pyogenes pSM19035 Addiction System
- Effects of Antineoplastic Agents on Cytoplasmic and Membrane-Bound Heat Shock Protein 70 (Hsp70) Levels
- Characterization of the VPS10 Domain of SorLA/LR11 as Binding Site for the Neuropeptide HA
- Substrate Specificity and Inhibitor Studies of a Membrane-Bound Ganglioside Sialidase Isolated from Human Brain Tissue
- Specificity Determinants in the Interaction of Apolipoprotein(a) Kringles with Tetranectin and LDL
- Amylin-Induced Cytotoxicity Is Associated with Activation of Caspase-3 and MAP Kinases
- 2-Alkyl-3-Alkylamino-2H-Benzo- and Pyridothiadiazine 1,1-Dioxides: From K+ATP Channel Openers to Ca++ Channel Blockers?
- Pre-Loading of Chlorophyll Synthase with Tetraprenyl Diphosphate Is an Obligatory Step in Chlorophyll Biosynthesis
- Molecular and Immunological Characterization of Profilin from Mugwort Pollen
- The Archaeal Respiratory Supercomplex SoxM from S.acidocaldarius Combines Features of Quinole and Cytochrome c Oxidases
- Benzodiazepine Binding Studies on Living Cells: Application of Small Ligands for Fluorescence Correlation Spectroscopy
- Individual Recombinant Thyroglobulin Type-1 Domains Are Substrates for Lysosomal Cysteine Proteinases
- Characterization of Legumain
- Molecular Cloning and Chimerisation of an Inhibitory Anti-Cathepsin B Antibody and Its Expression in Chinese Hamster Ovary Cells
- Application of Peptides Containing the Cleavage Sequence of Pro-TNFα in Assessing TACE Activity of Whole Cells
