Kinetics of the O-methylating system for isoprenaline in the trachea and aorta of rabbit.
Segments of tracheal smooth muscle or aorta from rabbits pretreated with reserpine (1 mg/kg) were incubated in 3H-isoprenaline (0.5-60 mumol/l). Steady-state rates of O-methylation were determined by measuring the formation of 3-O-methylisoprenaline (OMI) after incubation of tracheal and aortic tissues for 30 min and 10 min, respectively. The steady-state O-methylation of isoprenaline in rabbit trachea was saturable, at least up to 60 mumol/l isoprenaline. In rabbit aorta, the O-methylation appeared to be saturable up to 30 mumol/l isoprenaline, but the rate of O-methylation increased for higher concentrations. The Km values for the saturable component of O-methylation were 11.8 mumol/l in trachea and 3.03 mumol/l in aorta. The Vmax values were 0.51 nmol X g-1 X min-1 in trachea and 0.56 nmol X g-1 X min-1 in aorta. In tissues incubated in 0.5 mumol/l isoprenaline, 100 mumol/l corticosterone caused 78% inhibition of OMI formation in trachea and 86% inhibition in aorta. There was no inhibition of OMI formation by 100 mumol/l corticosterone in tracheal or aortic tissues incubated in 60 mumol/l isoprenaline. Model calculations showed that the experimental results in trachea and aorta (3. above) were consistent with (a) entry of isoprenaline into the cells in the tissues by extraneuronal uptake and diffusion, and (b) exposure of the isoprenaline to intracellular catechol-O-methyltransferase with Vmax enzyme much less than Vmax uptake.
D000818:Animals; D001011:Aorta / Q000378:metabolism; D002394:Catechol O-Methyltransferase / Q000378:metabolism; D003345:Corticosterone / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008745:Methylation; D008954:Models, Biological; D009130:Muscle, Smooth / Q000378:metabolism*; D011817:Rabbits; D012110:Reserpine / Q000494:pharmacology; D014132:Trachea / Q000378:metabolism
D012110:Reserpine; D002394:Catechol O-Methyltransferase; D007545:Isoproterenol; D003345:Corticosterone