(1) The renin-angiotensinogen reaction in the rat was examined in detail under various conditions of pH and temperature. Angiotensinase activity in the kidney extract and the plasma was also determined.
(2) pH-activity curves of angiotensinases in the plasma, red cells, and kidney extracts were determined. Angiotensinases in the kidney tissue had different pH-characteristics from those of the plasma or red cells.
(3) Incubation of the aid-treated extract with the heparinized plasma resulted in the maximum formation of angiotensin at pH 6.5, regardless of the presence of EDTA.
(4) At pH 5.5 formation of angiotensin continued to increase until 40min., and then formed a plateau, by incubating the acid- and alkaline-treated extract of 1/30 concentration at 37°C.
(5) At pH 6.5 formation of angiotensin reached the maximum after 20 to 40min., and then decreased slightly, by incubating the acid- treated extract of 1/30 concentration at 37°C with 3.3×10
-3 M EDTA.
(6) At pH 8.0 incubation of the acid-treated extract at 20°C resulted in a linear increase of angiotensin formation until 80min.
(7) Formation of angiotensin with various concentrations of the kidney extracts was determined under the above 3 conditions, where the effect of angiotensinases was relatively low and formation of angiotensin increased with the incubation time for a sufficient length.
(8) The reaction constants (K) calculated from the above results had a linear relationship with a wide range of concentrations of kidney extract at pH 8.0.
(9) Incubation of the acid-treated kidney extract with the plasma at pH 8.0 and 20°C for 30min. gave a more reliable method for the determination of renin in rat kidney.
View full abstract