Mutagenesis of the borage Delta(6) fatty acid desaturase.
The consensus sequence of the third histidine box of a range of Delta(5), Delta(6), Delta(8) and sphingolipid desaturases differs from that of the membrane-bound non-fusion Delta(12) and Delta(15) desaturases in the presence of glutamine instead of histidine. We have used site-directed mutagenesis to determine the importance of glutamine and other residues of the third histidine box and created a chimaeric enzyme to determine the ability of the Cyt b(5) fusion domain from the plant sphingolipid desaturase to substitute for the endogenous domain of the Delta(6) desaturase.
Sayanova
O
O
IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol BS41 9AF, UK. olga.sayanova@bbsrc.ac.uk
Beaudoin
F
F
Libisch
B
B
Shewry
P
P
Napier
J
J
D019943:Amino Acid Substitution; D017360:Arabidopsis / Q000201:enzymology; D019659:Asteraceae / Q000201:enzymology*; D015786:Cytochromes b5 / Q000737:chemistry / Q000378:metabolism; D044943:Fatty Acid Desaturases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D050602:Linoleoyl-CoA Desaturase; D016297:Mutagenesis, Site-Directed; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D014675:Vegetables / Q000201:enzymology
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D006639:Histidine; D015786:Cytochromes b5; D044943:Fatty Acid Desaturases; D050602:Linoleoyl-CoA Desaturase; C119257:delta-8 fatty acid desaturase