Functional identification of AtFao3, a membrane bound long chain alcohol oxidase in Arabidopsis thaliana.
The Arabidopsis thalina genome database was searched for homologues of the Candida cloacae fao1 gene which encodes a membrane bound, flavin-containing, hydrogen peroxide generating, long chain alcohol oxidase. This gene has not been isolated from plants or animals. Four putative candidates were found in the database but their function has not been proven. The cDNAs for two of them were cloned by RT-PCR from Arabidopis suspension culture and one of them [AtFAO3] was overexpressed in Escherichia coli and shown to functionally express long chain alcohol oxidase activity. The protein has been solubilised and retains biological activity thereby preparing the way for crystallographic studies. This is the first functional proof identifying a long chain alcohol oxidase in higher plants.
Cheng
Qi
Q
Department of Biological Sciences, University of Durham, South Road, Durham DH1 3LE, UK.
Liu
Huan-Ting
HT
Bombelli
Paolo
P
Smith
Alison
A
Slabas
Antoni R
AR
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001483:Base Sequence; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase