Document Detail


A novel pathway for transport and metabolism of a fluorescent phosphatidic acid analog in yeast.
MedLine Citation:
PMID:  11208128     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Phosphatidic acid is a central intermediate of biosynthetic lipid metabolism as well as an important signaling molecule in the cell. These studies assess the internalization, or retrograde transport, and metabolism of phosphatidic acid in yeast using a fluorescent analog. An analog of phosphatidic acid fluorescently labeled at the sn-2 position with N-4-nitrobenz-2-oxa-1, 3-diazole-aminocaproic acid (NBD-phosphatidic acid) was introduced to yeast cells by spontaneous transfer from phospholipid vesicles. Transport and metabolism of the NBD-phosphatidic acid were then monitored by fluorescence spectrophotometry, fluorescence microscopy and routine biochemical methods. Primary metabolites of the NBD-phosphatidic acid in yeast were found to be NBD-diacylgycerol and NBD-phosphatidylinositol. Experiments in cells possessing different levels of phosphatidate phosphatase activity suggest that conversion of the NBD-phosphatidic acid to NBD-diacylglycerol is not a pre-requisite for internalization in yeast. Internalization is sensitive to decreased temperature, but neither ATP depletion nor a sec6-4 mutation, which interrupts endocytosis, has an affect. Thus, internalization of NBD-phosphatidic acid apparently occurs via a non-endocytic route. These characteristics of retrograde transport of NBD-phosphatidic acid in yeast differ significantly from transport of other NBD-phospholipids in yeast as well as NBD-phosphatidic acid transport in mammalian fibroblasts.
Authors:
P J Trotter
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Traffic (Copenhagen, Denmark)     Volume:  1     ISSN:  1398-9219     ISO Abbreviation:  Traffic     Publication Date:  2000 May 
Date Detail:
Created Date:  2001-03-20     Completed Date:  2001-04-12     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  100939340     Medline TA:  Traffic     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  425-34     Citation Subset:  IM    
Affiliation:
Institute for Cellular and Molecular Biology and Division of Nutritional Sciences, University of Texas, Austin, TX 78712, USA. trotter@mail.utexas.edu
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MeSH Terms
Descriptor/Qualifier:
4-Chloro-7-nitrobenzofurazan / analogs & derivatives*,  metabolism*
Adenosine Triphosphate / metabolism
Biological Transport, Active
Carrier Proteins / genetics
Diglycerides / metabolism
Energy Metabolism
Fluorescent Dyes / metabolism*
Genes, Fungal
Kinetics
Models, Biological
Phosphatidic Acids / metabolism*
Saccharomyces cerevisiae / genetics,  metabolism*
Saccharomyces cerevisiae Proteins*
Temperature
Vesicular Transport Proteins
Chemical
Reg. No./Substance:
0/1-oleoyl-2-((N-(7-(nitro)-2,1,3-benzoxadiazol-4-yl)amino)hexanoyl)glycero-3-phosphate; 0/Carrier Proteins; 0/Diglycerides; 0/Fluorescent Dyes; 0/Phosphatidic Acids; 0/SEC6 protein, S cerevisiae; 0/Saccharomyces cerevisiae Proteins; 0/Vesicular Transport Proteins; 10199-89-0/4-Chloro-7-nitrobenzofurazan; 155581-64-9/1-oleoyl-2-hexanoyl-NBD-glycerol; 56-65-5/Adenosine Triphosphate

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


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