Identification of the active site of human renin with use of new fluorogenic peptides.

Fluorogenic peptide substrates were synthesized in which amino acid residues corresponded to the C-terminal and the N-terminal sides of the site of human angiotensinogen cleaved by renin. Compared with the synthetic substrates of renin previously reported, these fluorogenic substrates had practical advantages in that their digestion products could be rapidly separated and sensitively detected by high-pressure liquid chromatography with a fluorescence detector. The recombinant human renin and human plasma split Leu-Val, which cleavage site is similar to that in human angiotensinogen. The kinetic parameters of the reaction of renin using these substrates were calculated. There seemed to be at least eight subsites in the active site of recombinant human renin, to judge from the enzyme-substrate binding characteristics. The two histidine residues (S5 and S'3) in the octapeptide His-Pro-Phe-His-Leu-Val-Ile-His were important in the enzyme action.