PDBsum entry 1t32

Hydrolase

PDB id: 1t32

Contents

Protein chain

224 a.a. *

Ligands

SO4 ×5

OHH

Waters ×261

* Residue conservation analysis

PDB id:

1t32

Name:

Hydrolase

Title:

A dual inhibitor of the leukocyte proteases cathepsin g and chymase with therapeutic efficacy in animals models of inflammation

Structure:

Cathepsin g. Chain: a. Synonym: cg. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: human

Resolution:

1.85Å R-factor: 0.172 R-free: 0.213

Authors:

L.De Garavilla,M.N.Greco,E.C.Giardino,G.I.Wells,B.J.Haertlein, J.A.Kauffman,T.W.Corcoran,C.K.Derian,A.J.Eckardt,W.M.Abraham, N.Sukumar,Z.Chen,A.O.Pineda,F.S.Mathews,E.Di Cera,P.Andrade-Gordon, B.P.Damiano,B.E.Maryanoff

Key ref:

L.de Garavilla et al. (2005). A novel, potent dual inhibitor of the leukocyte proteases cathepsin G and chymase: molecular mechanisms and anti-inflammatory activity in vivo. J Biol Chem, 280, 18001-18007. PubMed id: 15741158 DOI: 10.1074/jbc.M501302200

Date:

23-Apr-04 Release date: 01-Mar-05

Protein chain

P08311  (CATG_HUMAN) -  Cathepsin G from Homo sapiens

Seq: 255 a.a.

Struc: 224 a.a.

Enzyme reactions

• Enzyme class: E.C.3.4.21.20 - cathepsin G.
• Reaction: Specificity similar to chymotrypsin C.

DOI no: 10.1074/jbc.M501302200

J Biol Chem 280:18001-18007 (2005)

PubMed id: 15741158

A novel, potent dual inhibitor of the leukocyte proteases cathepsin G and chymase: molecular mechanisms and anti-inflammatory activity in vivo.

L.de Garavilla, M.N.Greco, N.Sukumar, Z.W.Chen, A.O.Pineda, F.S.Mathews, E.Di Cera, E.C.Giardino, G.I.Wells, B.J.Haertlein, J.A.Kauffman, T.W.Corcoran, C.K.Derian, A.J.Eckardt, B.P.Damiano, P.Andrade-Gordon, B.E.Maryanoff.

ABSTRACT

Certain leukocytes release serine proteases that sustain inflammatory processes and cause disease conditions, such as asthma and chronic obstructive pulmonary disease. We identified beta-ketophosphonate 1 (JNJ-10311795; RWJ-355871) as a novel, potent dual inhibitor of neutrophil cathepsin G (K(i) = 38 nm) and mast cell chymase (K(i) = 2.3 nm). The x-ray crystal structures of 1 complexed with human cathepsin G (1.85 A) and human chymase (1.90 A) reveal the molecular basis of the dual inhibition. Ligand 1 occupies the S(1) and S(2) subsites of cathepsin G and chymase similarly, with the 2-naphthyl in S(1), the 1-naphthyl in S(2), and the phosphonate group in a complex network of hydrogen bonds. Surprisingly, however, the carboxamido-N-(naphthalene-2-carboxyl)piperidine group is found to bind in two distinct conformations. In cathepsin G, this group occupies the hydrophobic S(3)/S(4) subsites, whereas in chymase, it does not; rather, it folds onto the 1-naphthyl group of the inhibitor itself. Compound 1 exhibited noteworthy anti-inflammatory activity in rats for glycogen-induced peritonitis and lipopolysaccharide-induced airway inflammation. In addition to a marked reduction in neutrophil influx, 1 reversed increases in inflammatory mediators interleukin-1alpha, interleukin-1beta, tissue necrosis factor-alpha, and monocyte chemotactic protein-1 in the glycogen model and reversed increases in airway nitric oxide levels in the lipopolysaccharide model. These findings demonstrate that it is possible to inhibit both cathepsin G and chymase with a single molecule and suggest an exciting opportunity in the treatment of asthma and chronic obstructive pulmonary disease.

Selected figure(s)

Figure 2.
FIG. 2. X-ray crystallographic results. A, a view of the x-ray structure of 1 (yellow, with standard coloring for oxygen and nitrogen atoms) in the active site of Cat G, represented as a Connolly surface (green), at 1.85 Å resolution. B, a view of the x-ray structure of 1 (yellow, with standard coloring for oxygen and nitrogen atoms) in the active site of chymase, represented as a Connolly surface (green), at 1.9 Å resolution. C, a schematic representation of the interactions surrounding the S[1] and S[2] subsites for 1·Cat G. D, a schematic representation of the interactions surrounding the S[1] and S[2] subsites for 1·chymase. For A and B, certain amino acid residues and enzyme subsites are labeled. For C and D, ligand 1 is shown in its approximate orientation in each complex, with the amide bond that rotates 180° highlighted in red. The hydrogen bonds are depicted by dashed lines accompanied by the distances between the heavy atoms.

Figure 5.
FIG. 5. Inhibition of LPS-induced neutrophilia in the lungs of rats. Data are given for counts of white blood cells: total white blood cells (Total WBC), neutrophils (NEUT), lymphocytes (LYMPH), monocytes (MONO), eosinophils (EOS), and basophils (BASO). Each set of three clustered bars represents, from left to right, saline control (white), LPS treatment (black), and treatment with LPS + 1 (gray). The inset graph shows an expansion of the cell counts for monocytes, eosinophils, and basophils. Data are reported as mean ± S.E.; an asterisk indicates statistical significance at p 0.01 relative to the vehicle-treated LPS group (black bars).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2005, 280, 18001-18007) copyright 2005.

Figures were selected by the author.