Phase II biotransformation of carcinogens/atherogens in cultured aortic tissues and cells. I. Sulfation of 3-hydroxy-benzo(a)pyrene.
The capacity of polynuclear aromatic hydrocarbons to elicit profound effects on the development of avian aortic atheromata has raised questions regarding the biotransformation of polynuclear aromatic hydrocarbons in the target (aortic) tissue. Results of this investigation demonstrate the capacity of aortic enzymes to affect the sulfoconjugation of 3-hydroxy-benzo(a)pyrene and describe several characteristics of the aortic sulfotransferase activity. Conjugating activities measured in avian aortic tissues were approximately 10-20% of those assayed in corresponding preparations of avian hepatic tissues under the same reaction conditions. Activities were measured in homogenates, in a series of homogenate subfractions, in whole organ cultures, in cultured aortic endothelial cells, and in cultured aortic smooth muscle cell preparations. Sulfoconjugation was localized in the cytosolic fraction and kinetics in this fraction yielded a range of apparent Km values from 9 to 16 microM (mean = 11.8 +/- 3.1, n = 4) and a range of apparent Vmax values from 281 to 457 pmol/mg of protein/30 min (mean = 360 +/- 49, n = 4). Abdominal and thoracic segments of the aorta exhibited virtually identical specific activities. Also, activities assayed in cultured aortic smooth muscle cells were similar to those measured in cultured aortic endothelial cells. Capacity to generate adenosine 3'-phosphate 5'-phosphosulfate (PAPS) appeared to limit the reaction rate as judged by comparative investigations with PAPS and a PAPS-generating system. Aortic sulfatases actively hydrolyzed benzo(a)pyrene-3-O-sulfate. The sulfatase activity appeared to partially mask sulfotransferase activities measured in organ and cell culture preparations and in particulate subfractions of cellular homogenates.(ABSTRACT TRUNCATED AT 250 WORDS)
Yang
H Y
HY
Namkung
M J
MJ
Nelson
W L
WL
Juchau
M R
MR
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000201:enzymology* / Q000648:ultrastructure; D001161:Arteriosclerosis / Q000139:chemically induced / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D001580:Benzopyrenes / Q000378:metabolism*; D001711:Biotransformation; D002273:Carcinogens / Q000378:metabolism*; D002458:Cell Fractionation; D002645:Chickens; D046508:Culture Techniques; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure; D008297:Male; D008861:Microsomes / Q000201:enzymology; D009130:Muscle, Smooth / Q000201:enzymology; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism; D010920:Placenta / Q000166:cytology / Q000378:metabolism; D011083:Polycyclic Compounds / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013429:Sulfatases / Q000378:metabolism / Q000502:physiology
D001580:Benzopyrenes; D002273:Carcinogens; D011083:Polycyclic Compounds; D010724:Phosphoadenosine Phosphosulfate; C021338:3-hydroxybenzo(a)pyrene; D013429:Sulfatases