Document Detail


Sterol carrier protein-2 expression alters sphingolipid metabolism in transfected mouse L-cell fibroblasts.
MedLine Citation:
PMID:  16444586     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The influence of sterol carrier protein-2 (SCP-2) on the cellular metabolism of sphingolipids was examined in control mouse L-cells and stably transfected clones expressing the protein SCP-2. Three approaches were used to examine for differences; (1) compositional analysis of endogenous sphingolipid classes, (2) metabolism of NBD-ceramide, and (3) live cell labelling via endocytic uptake of BODIPY-sphingomyelin. SCP-2 over expression significantly altered the endogenous levels of both neutral and acidic sphingolipid classes. Among the neutral sphingolipids, expression of SCP-2 induced a 1.7-fold increase in the level of lactosylceramide (LacCer, p < 0.05) and a similar fold decrease in the level of the higher-order neutral glycosylceramides (p < 0.05). Among the acidic sphingolipids, SCP-2 resulted in a 5.2-fold decrease in the endogenous plasma membrane level of ganglioside GM1 (p < 0.03). Incubation of both control and transfected cell lines with NBD-ceramide resulted in the rapid establishment of a steady-state distribution of NBD-labelled sphingomyelin (NBD-SM) and glucosylceramide (NBD-GlcCer). In the SCP-2 expressing clones the conversion of NBD-Cer to NBD-GlcCer was 30% lower during incubation periods between 5 and 30 min (p < 0.025). Inspection of the cells by fluorescence microscopy after incubation with BODIPY labelled sphingomyelin (BODIPY-SM) revealed similar punctuated patterns with no distinguishable differences between the cell types. These results imply that SCP-2 plays a role in modulating enzymatic steps involved in metabolism of sphingolipid homeostasis.
Authors:
Daniel G Milis; Messiah K Moore; Barbara P Atshaves; Friedhelm Schroeder; John R Jefferson
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Molecular and cellular biochemistry     Volume:  283     ISSN:  0300-8177     ISO Abbreviation:  Mol. Cell. Biochem.     Publication Date:  2006 Feb 
Date Detail:
Created Date:  2006-01-30     Completed Date:  2006-04-18     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0364456     Medline TA:  Mol Cell Biochem     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  57-66     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Luther College, Decorah, Iowa, 52101-1045, USA.
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MeSH Terms
Descriptor/Qualifier:
4-Chloro-7-nitrobenzofurazan / analogs & derivatives,  pharmacokinetics,  pharmacology
Animals
Biological Transport
Carrier Proteins / metabolism*
Cell Membrane / metabolism
Ceramides / pharmacology
Fibroblasts / metabolism*
G(M1) Ganglioside / analogs & derivatives,  metabolism
Glucosylceramides / pharmacokinetics
Homeostasis
L Cells (Cell Line)
Mice
Microscopy, Fluorescence
Sphingolipids / metabolism*
Sphingomyelins / pharmacokinetics
Transfection
Grant Support
ID/Acronym/Agency:
GM31651/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Carrier Proteins; 0/Ceramides; 0/Glucosylceramides; 0/N-4-nitrobenzo-2-oxa-1,3-diazoleaminocaproyl sphingosylphosphorylcholine; 0/Sphingolipids; 0/Sphingomyelins; 0/ganglioside GM1alpha; 0/sterol carrier proteins; 10199-89-0/4-Chloro-7-nitrobenzofurazan; 37758-47-7/G(M1) Ganglioside; 86701-10-2/N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl sphingosine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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