| Biochemical and immunological studies on two distinct ganglioside-hydrolyzing sialidases from the particulate fraction of rat brain. | |
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MedLine Citation:
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PMID: 2398042 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Ganglioside-hydrolyzing sialidase activity was solubilized from rat brain particulate fraction by using Triton X-100 plus sodium deoxycholate. When chromatographed on AH-Sepharose 4B, the solubilized activity was resolved into two peaks, which were designated sialidases I and II in order of elution. The two sialidases were purified by using sequential chromatographies on Octyl-Sepharose CL-4B, Phenyl-Sepharose CL-4B, and Sephadex G-200. Sialidase II was purified further by Mono Q-FPLC. Overall purification was 450- and 2,150-fold, for sialidases I and II, respectively. Purified sialidases I and II were maximally active at near pH 5.0 and exhibited M = 70,000 by gel filtration. Sialidase I hydrolyzed gangliosides but scarcely other substrates including 4-methylumbelliferyl-NeuAc (4MU-NeuAc). Sialidase II hydrolyzed oligosaccharides, glycoproteins, and 4MU-NeuAc although gangliosides appeared to be preferential substrates. Sialidase II cleaved GM2 much faster than sialidase I. An antibody raised in rabbits against sialidase I reacted with only sialidase I and an antibody against sialidase II reacted with only sialidase II. A subcellular distribution study suggested sialidase I in the synaptosomal membrane and sialidase II in the synaptosomal and lysosomal membranes, and this was verified by using the above antibodies. |
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Authors:
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T Miyagi; J Sagawa; K Konno; S Handa; S Tsuiki |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of biochemistry Volume: 107 ISSN: 0021-924X ISO Abbreviation: J. Biochem. Publication Date: 1990 May |
Date Detail:
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Created Date: 1990-10-16 Completed Date: 1990-10-16 Revised Date: 2007-12-19 |
Medline Journal Info:
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Nlm Unique ID: 0376600 Medline TA: J Biochem Country: JAPAN |
Other Details:
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Languages: eng Pagination: 787-93 Citation Subset: IM |
Affiliation:
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Biochemistry Laboratory, Tohoku University, Miyagi. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies / immunology Brain / enzymology* Cell Membrane / enzymology Chromatography Hydrolysis Male Neuraminidase / immunology, metabolism* Organ Specificity Precipitin Tests Rats Rats, Inbred Strains Solubility Subcellular Fractions / enzymology* Substrate Specificity |
| Chemical | |
Reg. No./Substance:
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0/Antibodies; EC 3.2.1.18/GM3 ganglioside sialidase; EC 3.2.1.18/Neuraminidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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