Radiolabelled substrates for angiotensin converting enzyme.
Six [3H]benzoyl-tripeptides were prepared and tested as substrates for angiotensin converting enzyme. Each was prepared first as its [4-iodo]-benzoyl-analog, and an atom of 3H per molecule was introduced by catalytic dehalogenation in 3H2-gas. Kinetic parameters were measured at 37 degrees C using as buffer 0.05 M Hepes, pH 8.0 containing 0.1 M NaCl and 0.6 M Na2SO4. When the substrates were used at concentrations far below their respective Km values, fractional rates of substrate utilization per unit time for constant enzyme concentration were direct function of respective second order rate constants (Kc/Km). Although absolute values of Kc/Km differed for human enzyme as opposed to rabbit enzyme, relative values of Kc/Km were virtually identical. Similarly, relative rates of substrates utilization during passage through lungs of anesthetized rats were similar to relative values of Kc/Km measured in vitro. Thus, there is now a range of ACE substrates usable, in vitro and in vivo, under conditions of first order enzyme kinetics, conditions under which values of V/Km and Ki can be measured directly.
D000806:Angiotensin-Converting Enzyme Inhibitors; D000818:Animals; D006801:Humans; D007700:Kinetics; D008168:Lung / Q000201:enzymology; D009842:Oligopeptides / Q000378:metabolism*; D007703:Peptidyl-Dipeptidase A / Q000097:blood; D011817:Rabbits; D011865:Radioisotope Dilution Technique; D051381:Rats; D013379:Substrate Specificity; D014316:Tritium
D000806:Angiotensin-Converting Enzyme Inhibitors; D009842:Oligopeptides; D014316:Tritium; D007703:Peptidyl-Dipeptidase A