Reference "Cloning and functional expression in yeast of a cDNA coding for an obtusifoliol 14alpha-demethylase (CYP51) in wheat"

Reference ID

9087

Title

Cloning and functional expression in yeast of a cDNA coding for an obtusifoliol 14alpha-demethylase (CYP51) in wheat

Source

Biochemical and biophysical research communications, 1997, vol. 230, pp. 381-385

Authors (10)

Cabello-Hurtado-F	Zimmerlin-A
Rahier-A	Taton-M
DeRose-R	Nedelkina-S
Batard-Y	Durst-F
Pallett-K-E	Werck-Reichhart-D

Abstract

Screening of a wheat cDNA library with an heterologous CYP81B1 probe from
Helianthus tuberosus led to the isolation of a partial cDNA coding a protein
with all the characteristics of a typical P450 with high homology (32-39%
identity) to the fungal and mammalian CYP51s. Extensive screening of several
wheat cDNA libraries isolated a longer cDNA (W516) coding a peptide of 453 amino
acids. Alignment of W516 with other P450 sequences revealed that it was missing
a segment corresponding to the N-terminal membrane anchor of the protein. The
corresponding segment from the yeast lanosterol 14alpha-demethylase was linked
to the partial wheat cDNA and the chimera expressed in Saccharomyces cerevisiae.
Compared to microsomes from control yeasts, membranes of yeast expressing the
chimera catalysed 14alpha-demethylation of obtusifoliol with an increased
efficiency relative to lanosterol demethylase activity. W516 is thus a plant
member of the most ancient and conserved P450 family, CYP51.