PDBsum entry 4lsz

Hydrolase

PDB id: 4lsz

Contents

Protein chains

140 a.a.

94 a.a.

153 a.a.

Waters ×141

PDB id:

4lsz

Name:

Hydrolase

Title:

Caspase-7 in complex with darpin d7.18

Structure:

Caspase-7 subunit p20. Chain: a, c. Fragment: caspase-7 subunit p20, unp residues 24-198. Synonym: casp-7, apoptotic protease mch-3, cmh-1, ice-like apoptotic protease 3, ice-lap3, caspase-7 subunit p20, caspase-7 subunit p11. Engineered: yes. Caspase-7 subunit p10. Chain: b, d. Fragment: caspase-7 subunit p10, unp residues 207-303.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: casp7, mch3. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic construct. Organism_taxid: 32630. Expression_system_taxid: 562

Resolution:

2.26Å R-factor: 0.176 R-free: 0.208

Authors:

A.Fluetsch,M.Lukarska,M.G.Gruetter

Key ref:

A.Flütsch et al. (2014). Combined inhibition of caspase 3 and caspase 7 by two highly selective DARPins slows down cellular demise. Biochem J, 461, 279-290. PubMed id: 24779913 DOI: 10.1042/BJ20131456

Date:

23-Jul-13 Release date: 02-Jul-14

Protein chains

P55210  (CASP7_HUMAN) -  Caspase-7 from Homo sapiens

Seq: 303 a.a.

Struc: 140 a.a.

Protein chains

P55210  (CASP7_HUMAN) -  Caspase-7 from Homo sapiens

Seq: 303 a.a.

Struc: 94 a.a.

Protein chains

No UniProt id for this chain

Struc: 153 a.a.

Enzyme reactions

• Enzyme class: Chains A, B, C, D: E.C.3.4.22.60 - caspase-7.

DOI no: 10.1042/BJ20131456

Biochem J 461:279-290 (2014)

PubMed id: 24779913

Combined inhibition of caspase 3 and caspase 7 by two highly selective DARPins slows down cellular demise.

A.Flütsch, R.Ackermann, T.Schroeder, M.Lukarska, G.J.Hausammann, C.Weinert, C.Briand, M.G.Grütter.

ABSTRACT

Caspases play important roles during apoptosis, inflammation and proliferation. The high homology among family members makes selective targeting of individual caspases difficult, which is necessary to precisely define the role of these enzymes. We have selected caspase-7-specific binders from a library of DARPins (designed ankyrin repeat proteins). The DARPins D7.18 and D7.43 bind specifically to procaspase 7 and active caspase 7, but not to other members of the family. Binding of the DARPins does not affect the active enzyme, but interferes with its activation by other caspases. The crystal structure of the caspase 7-D7.18 complex elucidates the high selectivity and the mode of inhibition. Combining these caspase-7-specific DARPins with the previously reported caspase-3-inhibitory DARPin D3.4S76R reduces the activity of caspase 3 and 7 in double-transfected HeLa cells during apoptosis. In addition, these cells showed less susceptibility to TRAIL (tumour-necrosis-factor-related apoptosis-inducing ligand)-induced apoptosis in living cell experiments. D7.18 and D7.43 are therefore novel tools for in vitro studies on procaspase 7 activation as well as for clarifying the role of its activation in different cellular processes. If applied in combination with D3.4S76R, they represent an excellent instrument to increase our understanding of these enzymes during various cellular processes.