Caspase-3/Sandbox
| |||||||||
| Crystal Structure of Unliganded Human Caspase-3 | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Gene: | CASP3 OR CPP32 (Homo sapiens) | ||||||||
| |||||||||
| |||||||||
| Resources: | FirstGlance, OCA, PDBsum, RCSB | ||||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||||
Structure & FunctionStructure & Function
StructureStructure
(Structural Description) Caspase-3 chains are classified as alpha/beta, with one chain containing a 3-layer(aba) sandwich with Rossmann fold topology (This fold is made up of three or more parallel beta strands linked by two alpha helices in the topological order beta-alpha-beta-alpha-beta) and another chain containing a 2-layer sandwich with alpha-beta plaits.
The active form is made up of two chains, of which can be kept in its inactive form by inhibitors such as DEVD-CHO seen in 1pau.
DomainsDomains
Folds and MotifsFolds and Motifs
Posttranslational ModificationsPosttranslational Modifications
Active Site/Ligand BindingActive Site/Ligand Binding
This is a morph of caspase-3 from its uninhibited form to inhibition by novel irreversible inhibitor B92 ((3S)-3-({[(5S,10aS)-2-{(2S)-4-carboxy-2-[(phenylacetyl)amino]butyl}-1,3-dioxo-2,3,5,7,8,9,10,10a-octahydro-1H-[1,2,4]triazolo[1,2-a]cinnolin-5-yl]carbonyl}amino)-4-oxopentanoic acid). The involves, and . (Wang, Watt et al. 2010) When cleaved into its active form, caspase-3 is then able to bind to its substrates via recognition of DEVD consensus sequence. Caspase-3, using its active site cysteine residue, is then able to cleave the substrate at the Asp residue occupying the P4 portion of the active site. This is a (1qx3 to 2cjx)showing the binding of modified DEVD substrate (zDEVD-cmk) to the active site of caspase-3. 
|
| ||||||||||
FunctionFunction
(Structural insights into its function)
Caspases are proteases that function via a cysteine residue in the active site to cleave substrates after aspartic acid residues. Caspases are crucial for the initiation (e.g. caspase-8, -9, -10) and execution (e.g. caspase-3, -6, -7) of apoptosis, or programmed cell death either via the intrinsic or extrinsic pathway (Degterev, Boyce et al. 2003). In the intrinsic pathway, stimuli such as trophic factor withdrawal, UV irradiation, chemotherapeutics, DNA damage, endoplasmic reticulum (ER) stress, activates B cell lymphoma 2 (BCL-2) homology 3 (BH3)-only proteins like Bim or Bad, leading to BCL-2-associated X protein (BAX) and BCL-2 antagonist or killer (BAK) activation and mitochondrial outer membrane permeabilization (MOMP). Anti-apoptotic BCL-2 proteins prevent MOMP by binding BH3-only proteins and activated BAX or BAK. Following MOMP, release of various proteins from the mitochondrial intermembrane space (IMS) promotes caspase activation and apoptosis. Cytochrome c binds apoptotic protease-activating factor 1 (APAF1), inducing its oligomerization and thereby forming a structure termed the apoptosome that recruits and activates an initiator caspase, caspase-9. Caspase-9 cleaves and activates effector caspases, caspase-3 and caspase-7, leading to apoptosis through cleavage of death substrates such as Inhibitor of Caspase-activated Deoxyribonuclease (ICAD) and Poly ADP ribose polymerase (PARP). Mitochondrial release of second mitochondria-derived activator of caspase (SMAC; also known as DIABLO) and OMI (also known as HTRA2) neutralizes the caspase inhibitory function of X-linked inhibitor of apoptosis protein (XIAP). The extrinsic apoptotic pathway is initiated by the ligation of death receptors with their cognate ligands, leading to the recruitment of adaptor molecules such as Fas-associated death domain protein (FADD) and then caspase-8. This results in the dimerization and activation of caspase-8, which can then directly cleave and activate caspase-3 and caspase-7, leading to apoptosis. Crosstalk between the extrinsic and intrinsic pathways occurs through caspase-8 cleavage and activation of the BH3-only protein BH3-interacting domain death agonist (BID), the product of which (truncated BID; tBID) is required in some cell types for death receptor-induced apoptosis (Li, Zhu et al. 1998). It has become appreciated more recently that caspase activity, specifically caspase-8, is also required for T cell growth (Alam, Cohen et al. 1999; Kennedy, Kataoka et al. 1999; Misra, Jelley-Gibbs et al. 2005), and that the location and level of active caspases within cells may be a key determinant of survival or death (Misra, Russell et al. 2007; Koenig, Russell et al. 2008). We have previously observed that murine αβ T cells bearing high levels of caspase activity manifest increased rates of both cell growth and cell death (Dohrman, Russell et al. 2005).
Caspase-3Caspase-3
In its inactive form, procaspase-3 consist of a large subunit and small subunit, interjected by an aspartic acid residue. This aspartate is the site recognized by activated initiator caspases, such as caspase-8 and caspase-9. Upon cleavage, procaspase-3 is separated into two subunits, p18/20 and p12/10 respectively, which heterodimerize to yield the active form of caspase-3. In its active form, caspase-3 is able to cleave substrates such as ICAD (inhibitor of caspase-activated deoxyribonuclease). Cleavage of ICAD leads to abrogation of its inhibitory effect on CAD, allowing CAD to migrate into the nucleus and cause double-strand breaks in DNA, thus contributing to apoptosis(Enari, Sakahira et al. 1998; Sakahira, Enari et al. 1998).
DiseaseDisease
Although the role of caspases in chronic neurodegenerative disease is controversial, their role in acute neurodegenerative disease, such as nerve crush injury and stroke, may be more evident. Inhibition of caspase activity increases survival of neurons. Studies have also shown that caspases are downregulated to allow for unchecked survival of neoplastic cancers and autoimmune diseases. In particular, caspase-8 has been found to be silenced in neuroblastoma and humans with mutations in caspase-8 and -10 develop ALPS (autoimmune lymphoproliferative syndrome). These raise the possibility that caspase-3 may play a role as well. Knocking out caspase-3 in 129x1/SvJ mice leads to hydrocephalus and subsequently perinatal lethality. Caspase-3 knockout in C57BL/6J however appear to have no brain abnormalities.
(Fuentes-Prior and Salvesen, 2004)
Evolutionarily Related ProteinsEvolutionarily Related Proteins
To date, eighteen caspases have been identified. Caspases can largely be grouped into three subfamilies in humans: inflammatory (caspase-1, -4, and -5), effector (caspase-3, -6, and -7), and initiator caspases (-2, -8, -9, and -10). Caspase-11 and -12 substitutes for caspase-4 and -5, respectively, in mice. (Fuentes-Prior and Salvesen, 2004)
Solved StructuresSolved Structures
(Links to available structures) 3PCX Caspase-3 E246A, K242A Double Mutant http://www.pdb.org/pdb/explore/explore.do?structureId=3PCX 3PD1 Caspase-3 K242A http://www.pdb.org/pdb/explore/explore.do?structureId=3PD1 3PD0 Caspase-3 E246A http://www.pdb.org/pdb/explore/explore.do?structureId=3PD03KJF 3KJF Caspase-3 bound to a covalent inhibitor http://www.pdb.org/pdb/explore/explore.do?structureId=3KJF
3ITN
3H0E
3GJQ, 3GJR, 3GJS, 3GJT
3EDQ
3DEH, 3DEI, 3DEJ, 3DEK
2CNK, 2CNL, 2CNN, 2CNO, 2CDR
2J30, 2J31, 2J32, 2J33
2C1E, 2C2K, 2C2M, 2C2O
2H51
2H5J
2H65
2DKO, 2CJX, 2CJY
1RE1
1RHJ, 1RHK, 1RHM, 1RHQ, 1RHR, 1RHU
1NME, 1NMQ, 1NMS
1CP3
1PAU
1I3O
1QX3
References & NotesReferences & Notes
Alam, A., L. Y. Cohen, et al. (1999). "Early activation of caspases during T lymphocyte stimulation results in selective substrate cleavage in nonapoptotic cells." J Exp Med 190(12): 1879-90.
Aouad, S. M., L. Y. Cohen, et al. (2004). "Caspase-3 is a component of Fas death-inducing signaling complex in lipid rafts and its activity is required for complete caspase-8 activation during Fas-mediated cell death." J Immunol 172(4): 2316-23.
Budd, R. C. (2001). "Activation-induced cell death." Curr Opin Immunol 13(3): 356-62.
Degterev, A., M. Boyce, et al. (2003). "A decade of caspases." Oncogene 22(53): 8543-67.
Dohrman, A., J. Q. Russell, et al. (2005). "Cellular FLIP long form augments caspase activity and death of T cells through heterodimerization with and activation of caspase-8." J Immunol 175(1): 311-8.
Enari, M., H. Sakahira, et al. (1998). "A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD." Nature 391(6662): 43-50.
Kennedy, N. J., T. Kataoka, et al. (1999). "Caspase activation is required for T cell proliferation." J Exp Med 190(12): 1891-6.
Koenig, A., J. Q. Russell, et al. (2008). "Spatial differences in active caspase-8 defines its role in T-cell activation versus cell death." Cell Death Differ 15(11): 1701-11.
Li, H., H. Zhu, et al. (1998). "Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis." Cell 94(4): 491-501.
Misra, R. S., D. M. Jelley-Gibbs, et al. (2005). "Effector CD4+ T cells generate intermediate caspase activity and cleavage of caspase-8 substrates." J Immunol 174(7): 3999-4009.
Misra, R. S., J. Q. Russell, et al. (2007). "Caspase-8 and c-FLIPL associate in lipid rafts with NF-kappaB adaptors during T cell activation." J Biol Chem 282(27): 19365-74.
Sakahira, H., M. Enari, et al. (1998). "Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis." Nature 391(6662): 96-9.
Vincent, M. S., K. Roessner, et al. (1996). "Apoptosis of Fashigh CD4+ synovial T cells by borrelia-reactive Fas-ligand(high) gamma delta T cells in Lyme arthritis." J Exp Med 184(6): 2109-17.
Wang Z, Watt W, et al. (2010) Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors. Biochim Biophys Acta 1804(9):1817-31.
Wesselborg, S., O. Janssen, et al. (1993). "Induction of activation-driven death (apoptosis) in activated but not resting peripheral blood T cells." J Immunol 150(10): 4338-45.