KININ FORMATION BY CATHEPTIC ENZYME IN RAT STOMACH

Four uterine-contractile substances (a, b, c and d) were extracted from procedures with acetic acid, n-butanol, distilled water, and methanol from the reaction mixture obtained by incubating rat plasma kininogen with the kininforming enzyme in the rat stomach. Gradient and equilibrium chromatography on SP-Sephadex C-25 columns were carried out with the extract containing these materials (a, b, c and d). The materials could not be separated by chromatography on SP-Sephadex C-25, but were separated by high performance liquid chromatography (HPLC) with a Zorbax ODS reversed-phase column. All the materials (a, b, c and d) contracted the rat uterus and relaxed the rat duodenum. The uterine-contractile activity of the materials was abolished by chymotrypsin, but not by trypsin treatment. The retention times of materials a, c and d on HPLC were different from that of kallidin, MLBK, bradykinin (BK) and neurotensin; but the retention time of material b was equal to that of BK. The content ratio of a: b: c: d was 4: 4: 67: 25, calculated from the uterine-contractile activity of these materials. The apparent molecular weight of the major material c, estimated by gel chromatography, was 1650. Material c contracted the rat uterus (1.2×10−10 g/ml), relaxed the rat duodenum (2×10−10 g/ml), and produced a fall in rabbit blood pressure (4.4×10−8 g/kg). Material c was classified as a biologically BK-like peptide which is distinct from kallidin, MLBK, BK and neurotensin.