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PMID: 2006187 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Stearoyl-acyl-carrier-protein desaturase from higher plants is structurally unrelated to the animal and fungal homologs.

Shanklin J, Somerville C

Abstract

Stearoyl-acyl-carrier-protein (ACP) desaturase (EC 1.14.99.6) was purified to homogeneity from avocado mesocarp, and monospecific polyclonal antibodies directed against the protein were used to isolate full-length cDNA clones from Ricinus communis (castor) seed and Cucumis sativus (cucumber). The nucleotide sequence of the castor clone pRCD1 revealed an open reading frame of 1.2 kilobases encoding a 396-amino acid protein of 45 kDa. The cucumber clone pCSD1 encoded a homologous 396-amino acid protein with 88% amino acid identity to the castor clone. Expression of pRCD1 in Saccharomyces cerevisiae resulted in the accumulation of a functional stearoyl-ACP desaturase, demonstrating that the introduction of this single gene product was sufficient to confer soluble desaturase activity to yeast. There was no detectable identity between the deduced amino acid sequences of the castor delta 9-stearoyl-ACP desaturase and either the delta 9-stearoyl-CoA desaturase from rat or yeast or the delta 12 desaturase from Synechocystis, suggesting that these enzymes may have evolved independently. However, there was a 48-residue region of 29% amino acid sequence identity between residues 53 and 101 of the castor desaturase and the proximal border of the dehydratase region of the fatty acid synthase from yeast. Stearoyl-ACP mRNA was present at substantially higher levels in developing seeds than in leaf and root tissue, suggesting that expression of the delta 9 desaturase is developmentally regulated.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular DNA/genetics,isolation & purification Escherichia coli/genetics Mixed Function Oxygenases/genetics,isolation & purification,metabolism Molecular Sequence Data Open Reading Frames Plants/enzymology,genetics Rats Recombinant Proteins/isolation & purification,metabolism Saccharomyces cerevisiae/enzymology,genetics Sequence Homology, Nucleic Acid Species Specificity
Chemicals
Recombinant Proteins DNA Mixed Function Oxygenases acyl-(acyl-carrier-protein)desaturase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shanklin J
Department of Energy Plant Research Laboratory, Michigan State University, East Lansing 48824.
Somerville C
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18 references, click to expand
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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
1991-03-15
Pages
2510-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51262
Subset
IM
Databases
GENBANK
D90291, D90292, D90293, D90294, D90295, D90296, D90297, D90298, M59857, M59858, M62321
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