X-ray structures of five renin inhibitors bound to saccharopepsin: Exploration of active-site specificity

被引:9
作者
Cronin, NB
Badasso, MO
Tickle, IJ
Dreyer, T
Hoover, DJ
Rosati, RL
Humblet, CC
Lunney, EA
Cooper, JB [1 ]
机构
[1] Univ Southampton, Sch Biol Sci, Div Biochem & Mol Biol, Southampton SO16 7PX, Hants, England
[2] Warner Lambert Parke Davis, Parke Davis Pharmaceut Res, Dept Chem, Ann Arbor, MI 48106 USA
[3] Warner Lambert Parke Davis, Parke Davis Pharmaceut Res, Dept Pharmacol, Ann Arbor, MI 48106 USA
[4] Univ London Birkbeck Coll, Dept Crystallog, London WC1E 7HX, England
[5] Univ Minnesota, Dept Microbiol & Oral Sci, Minneapolis, MN 55455 USA
[6] Carlsberg Lab, Dept Chem, DK-2500 Copenhagen, Denmark
[7] Pfizer Inc, Div Cent Res, Dept Med Chem, Groton, CT 06340 USA
基金
英国生物技术与生命科学研究理事会;
关键词
renin inhibitors; aspartic proteinases; saccharopepsin;
D O I
10.1006/jmbi.2000.4181
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Saccharopepsin is a vacuolar aspartic proteinase involved in activation of a number of hydrolases. The enzyme has great structural homology to mammalian aspartic proteinases including human renin and we have used it as a model system to study the binding of renin inhibitors by X-ray crystallography. Five medium-to-high resolution structures of saccharopepsin complexed with transition-state analogue renin inhibitors were determined. The structure of a cyclic peptide inhibitor (PD-129,541) complexed with the proteinase was solved to 2.5 Angstrom, resolution. This inhibitor has low affinity for human renin yet binds very tightly to the yeast proteinase (K(i) = 4 nM). The high affinity of this inhibitor can be attributed to its bulky cyclic moiety spanning P(2)-P(3)' and other residues that appear to optimally fit the binding sub-sites of the enzyme. Superposition of the saccharopepsin structure on that of renin showed that a movement of the loop 286-301 relative to renin facilitates tighter binding of this inhibitor to saccharopepsin. Our 2.8 Angstrom resolution structure of the complex with CP-108,420 shows that its benzimidazole P(3) replacement retains one of the standard hydrogen bonds that normally involve the inhibitor's main-chain. This suggests a non-peptide lead in overcoming the problem of susceptible peptide bonds in the design of aspartic proteinase inhibitors. CP-72,647 which possesses a basic histidine residue at P(2), has a high affinity for renin (K(i) = 5 nM) but proves to be a poor inhibitor for saccharopepsin (K(i) = 3.7 muM) This may stem from the fact that the histidine residue would not bind favourably with the predominantly hydrophobic S(2) sub-site of saccharopepsin. (C) 2000 Academic Press.
引用
收藏
页码:745 / 760
页数:16
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