Analysis of non-heme iron in arachidonate 12-lipoxygenase of porcine leukocytes.
Arachidonate 12-lipoxygenase of porcine leukocytes, which was purified to homogeneity by immunoaffinity chromatography, was analyzed for iron content by atomic absorption spectrophotometry. The enzyme contained 0.70 +/- 0.09 g atom of iron per mol of enzyme (mean +/- S.D., n = 4). Inorganic iron, which was added to the enzyme solution as an internal standard, was recovered in almost 100% yield. Among various iron chelators tested, only 2,2'-dipyridyl at 1 mM inactivated the enzyme by 87%, but the enzyme was not reactivated by the addition of excess ferrous or ferric iron.
Matsuda
S
S
Department of Biochemistry, Tokushima University School of Medicine, Japan.
Suzuki
H
H
Yoshimoto
T
T
Yamamoto
S
S
Miyatake
A
A
D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D007501:Iron / Q000032:analysis*; D007962:Leukocytes / Q000201:enzymology*; D013054:Spectrophotometry, Atomic; D013552:Swine
D007501:Iron; D001092:Arachidonate 12-Lipoxygenase