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PMID: 8058731 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

The LCB2 gene of Saccharomyces and the related LCB1 gene encode subunits of serine palmitoyltransferase, the initial enzyme in sphingolipid synthesis.

Nagiec MM, Baltisberger JA, Wells GB, Lester RL, Dickson RC

Abstract

The first and committed step in synthesis of the ceramide moiety of sphingolipids is catalyzed by serine palmitoyltransferase (EC 2.3.1.50), which condenses palmitoyl-CoA and serine to form 3-ketosphinganine. This step is thought to be tightly regulated to control the synthesis of sphingolipids, but data supporting this hypothesis are lacking mainly because the enzyme has resisted purification and consequent characterization. Rather than attempting to purify the enzyme from normal cells, we have taken a different tack and opted to try and overproduce the enzyme to facilitate its purification. Here we demonstrate that overproduction in Saccharomyces cerevisiae requires expression of LCB1, a previously isolated yeast gene, and LCB2, the isolation and characterization of which we describe. Several lines of evidence argue that both genes encode subunits of the enzyme; however, biochemical evidence will be needed to substantiate this hypothesis. Although overproduction was modest, 2- to 4-fold, it should now be possible to devise improved overproduction vectors for yeast or other host organisms.

Related Genes
MeSH Terms
Acyltransferases/biosynthesis,genetics,metabolism Amino Acid Sequence Gene Expression Genes, Fungal Genotype Macromolecular Substances Molecular Sequence Data Plasmids Restriction Mapping Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Serine C-Palmitoyltransferase Sphingolipids/biosynthesis
Chemicals
Macromolecular Substances Saccharomyces cerevisiae Proteins Sphingolipids Acyltransferases LCB1 protein, S cerevisiae LCB2 protein, S cerevisiae Serine C-Palmitoyltransferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nagiec M M
Department of Biochemistry, University of Kentucky, Lexington 40536-0084.
Baltisberger J A
Wells G B
Lester R L
Dickson R C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1994-08-16
Pages
7899-902
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC44511
Subset
IM
Grants
NIGMS NIH HHS · GM41302 · United States
Databases
GENBANK
M95669
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