Identification of an essential sequence for dihydroceramide C-4 hydroxylase activity of mouse DES2.
Although the amino acid sequences of mouse DES1 (MDES1) and DES2 (MDES2) have 63% sequence identity, their enzymatic characteristics are quite different. MDES1 exhibits high dihydroceramide delta4-desaturase activity and very low C-4 hydroxylase activity, while MDES2 is similarly active as both a dihydroceramide delta4-desaturase and a C-4 hydroxylase. We constructed several chimeras of MDES1 and MDES2 and identified a region important for C-4 hydroxylase activity in MDES2. This region contains the sequence XAFGY (X=T or A or V; Y=T or N) and occurs on the C-terminal side of the first His-box of MDES2. We confirmed the conservation of this region in DES2 family members sequenced from humans, pigs, rats, chickens, zebrafish, and Xenopus.
Sphingolipid Expression Laboratory, Frontier Research System, RIKEN, 2-1, Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Fumio
F
Omae
Masao
M
Miyazaki
Ayako
A
Enomoto
Akemi
A
Suzuki
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D009097:Multienzyme Complexes; D011994:Recombinant Proteins; C489141:Degs2 protein, mouse; D010088:Oxidoreductases; C108138:dihydroceramide desaturase; D006867:Hydrolases