| The use of liposomes as acceptors for the assay of lipid glycosyltransferases from rat brain. | |
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MedLine Citation:
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PMID: 427178 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Preparation and characterization of sonicated vesicles of various lipid composition containing hydroxy and normal fatty acid ceramides are reported. Such vesicles have been successfully used for the first time as acceptors for the assays of lipid glycosyltransferases, UDP-galactose:ceramide galactosyltransferase and UDPglucose: ceramide glucosyltransferase. Stability of the vesicles and the optimal enzyme activities were the criteria used to select the final composition of the vesicles. The activities of the glycosyltransferases were dependent not only on the appropriate assay conditions but also on the type and source of the phospholipids used to form the liposomes. Ceramides containing normal fatty acids were incorporated into phosphatidylcholine vesicles in a molar ratio of 1 : 3.4 and used as the acceptor for the assay of UDPglucose:ceramide glucostyltransferase. For the assay UDP-galactose:ceramide galactosyltransferase, vesicles were prepared by sonication of bovine brain ethanolamine phospholipids, phosphatidylcholine and ceramide containing alpha-hydroxy fatty acids, in a molar ratio of 6 : 0.57 : 1. The size of the vesicles as determined by electron microscopic measurement ranged mostly between 200--500 A. The results obtained by selective labelling of the outer surface amino groups with the membrane-impermeable reagent, 2,4,6-trinitrobenzenesulfonic acid, indicated that the ethanolamine phospholipid-containing liposomes consisted of closed vesicles. After incubation with the appropriate cofactors and labelled sugar nucleotides, the radioactive reaction products were shown to cochromatograph with the authentic standards by thin-layer chromatography and autoradiography. |
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Authors:
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A Cestelli; F V White; E Costantino-Ceccarini |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biochimica et biophysica acta Volume: 572 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 1979 Feb |
Date Detail:
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Created Date: 1979-06-26 Completed Date: 1979-06-26 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 283-92 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Brain / enzymology* Ceramides Cerebrosides Galactosyltransferases / metabolism* Glucosyltransferases / metabolism* Kinetics Liposomes* Microsomes / enzymology Rats |
| Chemical | |
Reg. No./Substance:
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0/Ceramides; 0/Cerebrosides; 0/Liposomes; EC 2.4.1.-/Galactosyltransferases; EC 2.4.1.-/Glucosyltransferases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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