Immunological quantitation and immunohistochemical localization of leukotriene A4 hydrolase in guinea pig tissues.
We prepared a highly specific polyclonal antibody against leukotriene (LT) A4 hydrolase using a recombinant human enzyme. Using this antibody, we quantified LTA4 hydrolase protein content in the cytosols of guinea pig tissues. The enzyme protein content correlated well with the enzyme activity with a correlation coefficient of 0.87. However, the enzyme activity per mg of the enzyme in the cytosols was low, particularly in the liver and adrenal gland, compared with the specific activity of the purified enzyme. These observations suggest the presence of inhibitory substances and/or inactive enzymes in the cytosols of these tissues. To determine the cellular localization of LTA4 hydrolase in tissues other than blood cells, we carried out immunohistochemical examinations of guinea pig tissues. We identified epithelial cells in the tracheobronchial system and gastrointestinal tract, smooth muscle cells in the bronchi and aorta, vascular endothelial cells, and the intestinal plexus as novel cellular sources of the enzyme in the parenchyme of the tissue. Thus, LTA4 hydrolase was widely distributed in various types of parenchymal cells in the tissues, and this observation warrants further investigations on the biological activities of LTB4 in these cells and tissues.
Ohishi
N
N
Department of Physiological Chemistry and Nutrition, Faculty of Medicine, University of Tokyo, Japan.
Minami
M
M
Kobayashi
J
J
Seyama
Y
Y
Hata
J
J
Yotsumoto
H
H
Takaku
F
F
Shimizu
T
T
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000276:immunology; D005260:Female; D006168:Guinea Pigs; D007106:Immune Sera; D015151:Immunoblotting; D007150:Immunohistochemistry; D009928:Organ Specificity
D000906:Antibodies; D007106:Immune Sera; D004851:Epoxide Hydrolases; C043134:leukotriene A4 hydrolase