Molecular cloning and functional expression of rat leukotriene A4 hydrolase using the polymerase chain reaction.
We isolated a cDNA encoding rat leukotriene A4 (LTA4) hydrolase from mesangial cells by the polymerase chain reaction according to the human amino acid sequence. The deduced amino acid sequence shows that rat LTA4 hydrolase is a 609 amino acid protein with an Mr 69 kDa. Comparison of human LTA4 hydrolase revealed 93% homology, and include zinc-binding motifs of aminopeptidases. COS-7 cells transfected with the cDNA revealed substantial LTA4 hydrolase activity, and their activities were abolished by preincubation with captopril, representing the first reported cDNA expression of recombinant enzyme in mammalian cells. RNA blot analysis indicated that LTA4 hydrolase was expressed in glomerular endothelial, epithelial and mesangial cells.
Makita
N
N
Department of Medicine, Vanderbilt University, Nashville, TN 37232.
Funk
C D
CD
Imai
E
E
Hoover
R L
RL
Badr
K F
KF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004851:Epoxide Hydrolases / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D004851:Epoxide Hydrolases; C043134:leukotriene A4 hydrolase