Crystal structure of NS-134 in complex with bovine cathepsin B: a two-headed epoxysuccinyl inhibitor extends along the entire active-site cleft

Biochem J. 2004 Jul 15;381(Pt 2):511-7. doi: 10.1042/BJ20040237.

Abstract

The crystal structure of the inhibitor NS-134 in complex with bovine cathepsin B reveals that functional groups attached to both sides of the epoxysuccinyl reactive group bind to the part of active-site cleft as predicted. The -Leu-Pro-OH side binds to the primed binding sites interacting with the His110 and His111 residues with its C-terminal carboxy group, whereas the -Leu-Gly-Meu (-Leu-Gly-Gly-OMe) part (Meu, methoxycarbonylmethyl) binds along the non-primed binding sites. Comparison with the propeptide structures of cathepsins revealed that the binding of the latter part is least similar to the procathepsin B structure; this result, together with the two-residue shift in positioning of the Leu-Gly-Gly part, suggests that the propeptide structures of the cognate enzymes may not be the best starting point for the design of reverse binding inhibitors.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Binding Sites
  • Cathepsin B / antagonists & inhibitors
  • Cathepsin B / chemistry*
  • Cathepsin B / isolation & purification
  • Cattle
  • Crystallography, X-Ray / methods*
  • Cysteine Proteinase Inhibitors / chemistry
  • Dipeptides / chemistry
  • Epoxy Compounds / chemistry
  • Kidney / enzymology
  • Leucine / analogs & derivatives*
  • Leucine / chemistry
  • Molecular Conformation
  • Molecular Structure
  • Oligopeptides / chemical synthesis
  • Oligopeptides / chemistry*
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Quaternary
  • Pyridines / chemistry
  • Pyrroles / chemical synthesis
  • Pyrroles / chemistry*

Substances

  • 1-(2-((3-(1-(((methoxycarbonylmethyl-carbamoyl)-methyl)-carbamoyl)-3-methyl-butylcarbamoyl)-oxiranecarbonyl)-amino)-4-methyl-pentanoyl)-pyrrolidine-2-carboxylic acid
  • CLIK 148
  • Cysteine Proteinase Inhibitors
  • Dipeptides
  • Epoxy Compounds
  • Oligopeptides
  • Pyridines
  • Pyrroles
  • N-(3-ethoxycarbonyloxirane-2-carbonyl)-isoleucyl-proline
  • N-(3-propylcarbamoyloxirane-2-carbonyl)-isoleucyl-proline
  • Cathepsin B
  • Leucine
  • E 64

Associated data

  • PDB/1SP4