Premium
Cloning and characterization of the Dictyostelium discoideum cycloartenol synthase cDNA
Author(s) -
Godzina Sharotka M.,
Lovato Martha A.,
Meyer Michelle M.,
Foster Kimberly A.,
Wilson William K.,
Gu Wei,
Hostos Eugenio L.,
Matsuda Seiichi P. T.
Publication year - 2000
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-000-0520-3
Subject(s) - cycloartenol , lanosterol , dictyostelium discoideum , saccharomyces cerevisiae , biochemistry , biology , complementary dna , atp synthase , eukaryote , gene , sterol , genome , cholesterol
Cycloartenol synthase converts oxidosqualene to cycloartenol, the first carbocyclic intermediate en route to sterols in plants and many protists. Presented here is the first cycloartenol synthase gene identified from a protist, the cellular slime mold Dictyostelium discoideum . The cDNA encodes an 81‐kDa predicted protein 50–52% identical to known higher plant cycloartenol synthases and 40–49% identical to known lanosterol synthases from fungi and mammals. The encoded protein expressed in transgenic Saccharomyces cerevisiae converted synthetic oxidosqualene to cycloartenol in vitro . This product was characterized by 1 H and 13 C nuclear magnetic resonance and gas chromatography‐mass spectrometry. The predicted protein sequence diverges sufficiently from the known cycloartenol synthase sequences to dramatically reduce the number of residues that are candidates for the catalytic difference between cycloartenol and lanosterol formation.