PDBsum entry: 1gfw

Hydrolase

PDB id: 1gfw

Contents

Protein chains

145 a.a. *

93 a.a. *

Ligands

MSI

Waters ×45

* Residue conservation analysis

PDB id:

1gfw

Name:

Hydrolase

Title:

The 2.8 angstrom crystal structure of caspase-3 (apopain or cpp32)in complex with an isatin sulfonamide inhibitor.

Structure:

Caspase-3 (apopain, p20). Chain: a. Fragment: activated mature caspase-3 (p20) without pro- domain or linker (residues 29-175). Engineered: yes. Caspase-3 (apopain, p10). Chain: b. Fragment: activated mature caspase-3 (p10) without pro- domain or linker (residues 181-277).

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Other_details: human t-lymphocyte cell line jurkat.

Biol. unit:

Octamer (from PQS)

Resolution:

2.80Å R-factor: 0.200 R-free: 0.285

Authors:

N.O.Concha,C.A.Janson

Key ref:

D.Lee et al. (2000). Potent and selective nonpeptide inhibitors of caspases 3 and 7 inhibit apoptosis and maintain cell functionality. J Biol Chem, 275, 16007-16014. PubMed id: 10821855 DOI: 10.1074/jbc.275.21.16007

Date:

16-Jun-00 Release date: 23-Jun-00

Supersedes:

1qa8

Protein chain

P42574  (CASP3_HUMAN) -  Caspase-3

Seq: 277 a.a.

Struc: 145 a.a.

Protein chain

P42574  (CASP3_HUMAN) -  Caspase-3

Seq: 277 a.a.

Struc: 93 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: Chains A, B: E.C.3.4.22.56 - Caspase-3.

 Gene Ontology (GO) functional annotation 

• Cellular component: cytoplasm (1 term)
• Biological process: apoptosis (2 terms)
• Biochemical function: cysteine-type peptidase activity (2 terms)

DOI no: 10.1074/jbc.275.21.16007

J Biol Chem 275:16007-16014 (2000)

PubMed id: 10821855

Potent and selective nonpeptide inhibitors of caspases 3 and 7 inhibit apoptosis and maintain cell functionality.

D.Lee, S.A.Long, J.L.Adams, G.Chan, K.S.Vaidya, T.A.Francis, K.Kikly, J.D.Winkler, C.M.Sung, C.Debouck, S.Richardson, M.A.Levy, W.E.DeWolf, P.M.Keller, T.Tomaszek, M.S.Head, M.D.Ryan, R.C.Haltiwanger, P.H.Liang, C.A.Janson, P.J.McDevitt, K.Johanson, N.O.Concha, W.Chan, S.S.Abdel-Meguid, A.M.Badger, M.W.Lark, D.P.Nadeau, L.J.Suva, M.Gowen, M.E.Nuttall.

ABSTRACT

Caspases have been strongly implicated to play an essential role in apoptosis. A critical question regarding the role(s) of these proteases is whether selective inhibition of an effector caspase(s) will prevent cell death. We have identified potent and selective non-peptide inhibitors of the effector caspases 3 and 7. The inhibition of apoptosis and maintenance of cell functionality with a caspase 3/7-selective inhibitor is demonstrated for the first time, and suggests that targeting these two caspases alone is sufficient for blocking apoptosis. Furthermore, an x-ray co-crystal structure of the complex between recombinant human caspase 3 and an isatin sulfonamide inhibitor has been solved to 2.8-A resolution. In contrast to previously reported peptide-based caspase inhibitors, the isatin sulfonamides derive their selectivity for caspases 3 and 7 by interacting primarily with the S(2) subsite, and do not bind in the caspase primary aspartic acid binding pocket (S(1)). These inhibitors blocked apoptosis in murine bone marrow neutrophils and human chondrocytes. Furthermore, in camptothecin-induced chondrocyte apoptosis, cell functionality as measured by type II collagen promoter activity is maintained, an activity considered essential for cartilage homeostasis. These data suggest that inhibiting chondrocyte cell death with a caspase 3/7-selective inhibitor may provide a novel therapeutic approach for the prevention and treatment of osteoarthritis, or other disease states characterized by excessive apoptosis.

Selected figure(s)

Figure 1.
Fig. 1. Caspase inhibitors.

Figure 2.
Fig. 2. A, 2F[o] F[c] electron density map. Stereo view of the final 2F[o] F[c] electron density map around the inhibitor and water molecule contoured at 1 level. The map was computed using data between 6.0 and 2.8 Å and model phases. A covalent bond links the inhibitor C-3 atom to Cys163S , the pyrrolidine ring binds in the S[2] binding pocket, and the phenoxy ring occupies the shallow S[3] binding site. In this complex the S[1] pocket is occupied by Wat518. The figure was prepared with BOBSCRIPT (47) and Raster3D. B, crystal structure of enzyme/inhibitor 4 complex. Stereo view of the molecular surface representation of the caspase 3 active site in complex with isatin sulfonamide 4. The catalytic residues His121 and Cys163 are colored by atom, the S[1] pocket is the unoccupied region behind the isatin ring (shown in black), and the hydrophobic S[2] pocket formed by Tyr204-Phe^256-Trp206 (left to right) is shown in magenta. Carbon atoms of inhibitor 4 are gray; oxygen atoms are red, nitrogen atom is blue, and sulfur atom is yellow. The figure was generated using the program MOLMOL (48).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2000, 275, 16007-16014) copyright 2000.

Figures were selected by the author.