Substrate specificity and inhibitor structure-activity relationships of recombinant human renin: implications in the in vivo evaluation of renin inhibitors.
Homogeneous, active recombinant human renin obtained from Chinese hamster ovary (CHO) cells was characterized in vitro by (i) determination of its relative rates of hydrolysis of plasma angiotensinogens (ANGs) from human, monkey, baboon, rat, pig, rabbit, hamster, and dog and (ii) analysis of several synthetic ANG-based, inhibitors ranging in IC50 from 10(-10) to 10(-6) M. Comparison of the recombinant human renin with human kidney renin showed that these enzymes were indistinguishable from each other in terms of their plasma ANG specificities and inhibition by synthetic renin inhibitors. Porcine kidney renin was also characterized and shown to display plasma ANG hydrolysis profiles and inhibitor potencies that were markedly different from those of human renins. Finally, the results using the above plasma ANGs extend previous studies showing that the substrate specificity of human renin may be influenced by the amino acid residues at P2 (i.e., Ile, Val, or Tyr) and P3 (i.e., His or Tyr) sites. The relevance of these data to in vivo evaluation of renin inhibitors in animal models is discussed.
Evans
D B
DB
Biotechnology, Upjohn Company, Kalamazoo, Michigan 49001.
Cornette
J C
JC
Sawyer
T K
TK
Staples
D J
DJ
de Vaux
A E
AE
Sharma
S K
SK
D000595:Amino Acid Sequence; D000808:Angiotensinogen / Q000097:blood / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004285:Dogs; D006801:Humans; D006868:Hydrolysis; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012083:Renin / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013552:Swine; D000092502:Renin Inhibitors
D000808:Angiotensinogen; D011994:Recombinant Proteins; D012083:Renin; D000092502:Renin Inhibitors