Binding of Caspase-3 Prodomain to Heat Shock Protein 27 Regulates Monocyte Apoptosis by Inhibiting Caspase-3 Proteolytic Activation

Voß, Oliver; Batra, Sanjay; Kolattukudy, Sunny J.; González-Mejia, M. Elba; Smith, Jeffrey B.; Doseff, Andrea I.

doi:10.1074/jbc.m701740200

Cited by 152 publications

(151 citation statements)

References 36 publications

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“…For example, HSP27 can inhibit apoptosis by reducing caspase activity (33), decrease cytochrome c release from mitochondria (34), and/or inactivate the apoptosis signal-regulating kinase 1/JNK prodeath signaling cascade (17). A number of previous studies reported the neuroprotective effects of HSP27 against cerebral I/R injury when globally expressed in the brain (15)(16)(17)22).…”

Section: Discussionmentioning

confidence: 99%

Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood–brain barrier disruption after ischemic brain injury

Shi

Jiang

Zhang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

120

144

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The damage borne by the endothelial cells (ECs) forming the blood–brain barrier (BBB) during ischemic stroke and other neurological conditions disrupts the structure and function of the neurovascular unit and contributes to poor patient outcomes. We recently reported that structural aberrations in brain microvascular ECs—namely, uncontrolled actin polymerization and subsequent disassembly of junctional proteins, are a possible cause of the early onset BBB breach that arises within 30–60 min of reperfusion after transient focal ischemia. Here, we investigated the role of heat shock protein 27 (HSP27) as a direct inhibitor of actin polymerization and protectant against BBB disruption after ischemia/reperfusion (I/R). Using in vivo and in vitro models, we found that targeted overexpression of HSP27 specifically within ECs—but not within neurons—ameliorated BBB impairment 1–24 h after I/R. Mechanistically, HSP27 suppressed I/R-induced aberrant actin polymerization, stress fiber formation, and junctional protein translocation in brain microvascular ECs, independent of its protective actions against cell death. By preserving BBB integrity after I/R, EC-targeted HSP27 overexpression attenuated the infiltration of potentially destructive neutrophils and macrophages into brain parenchyma, thereby improving long-term stroke outcome. Notably, early poststroke administration of HSP27 attached to a cell-penetrating transduction domain (TAT-HSP27) rapidly elevated HSP27 levels in brain microvessels and ameliorated I/R-induced BBB disruption and subsequent neurological deficits. Thus, the present study demonstrates that HSP27 can function at the EC level to preserve BBB integrity after I/R brain injury. HSP27 may be a therapeutic agent for ischemic stroke and other neurological conditions involving BBB breakdown.

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Section: Discussionmentioning

confidence: 99%

Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood–brain barrier disruption after ischemic brain injury

Shi

Jiang

Zhang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

120

144

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“…42 However, other possible anti-apoptotic proteins could associate with the prodomain of caspase-6 in an analogous manner to the association of Hsp27 with the prodomain of caspase-3, which results in the inhibition of the autocatalytic processing of caspase-3 from its p20 to its p17 form that is required for its efficient processing and activation. 43 In summary, in contrast to the established model using dATP-activated cell lysates, caspase-7 in addition to caspase-3 can directly process caspase-6, and subsequently the processed caspase-6 can directly process caspase-8. Reagents.…”

Section: Discussionmentioning

confidence: 99%

Ordering of caspases in cells undergoing apoptosis by the intrinsic pathway

et al. 2009

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Caspases are a family of aspartate-specific cysteine proteases responsible for the biochemical and morphological changes that occur during the execution phase of apoptosis. The hierarchical ordering of caspases has been clearly established using dATPactivated cell lysates to model the intrinsic pathway induced by initial mitochondrial perturbation. In this model, caspase-9, the apical caspase, directly processes and activates the effector caspases, caspase-3 and -7, and then active caspase-3 but not caspase-7, processes caspase-2 and -6, and subsequently the activated caspase-6 processes caspase-8 and -10. To address the possibility that this model in vitro system might not reflect the precise ordering of caspases in intact cells, we have examined this possibility in cells induced to undergo apoptosis by activation of the intrinsic pathway. We have used caspase deficient cells, small interference RNA for caspase-6 and -7, and a specific caspase-3 inhibitor. In contrast to the earlier in vitro studies, we now show that in intact cells caspase-7 can also directly process and activate caspase-2 and -6. The processing of caspase-2 and -6 occurs within the cytoplasm and active caspase-6 is then responsible for both the processing of caspase-8 and the cleavage of caspase-6 substrates, including lamin A/C. Apoptosis, a major form of cell death used to remove unwanted or excess cells, is characterized by a plethora of morphological and biochemical changes resulting primarily from the activation of a family of cysteine proteases, which cleave their substrates at specific aspartate residues. 1-4 Two major apoptotic pathways have been described, the intrinsic pathway involving initial mitochondrial perturbation resulting from cellular stress or cytotoxic insults and the extrinsic pathway, which is triggered by the activation of death receptors of the TNF family. 5-7 Caspase-8 and -9, the apical caspases in the extrinsic and intrinsic pathways, possess a protein interaction prodomain, which result in their recruitment and subsequent activation within cellular complexes, namely the death-inducing signaling complex (DISC) and the Apaf-1 apoptosome, respectively, and the subsequent activation of effector caspases, such as caspase-3, -6 and -7. 5,[8][9][10] Caspases are synthesized as single-chain zymogens, containing an N-terminal prodomain, as well as large (B20 kDa) and small (B10 kDa) subunits, with the basic catalytic domain forming from a large and small subunits. 1,3,4 Caspase-2 also has a sizeable prodomain, but it is unclear whether it should be considered an initiator or an effector caspase. 3 Although caspase-2 has been proposed to act upstream of mitochondria and to be activated within the PIDDosome, it may also be activated downstream of caspase-3. 11-14 Using recombinant proteins, caspase-3, -8 and to a lesser extent caspase-7 can process caspase-2. 15 The physiological role of caspase-2 is not known and no marked phenotype is observed in caspase-2 À/À mice. Caspase-6 is generally considered to be an effector casp...

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“…Procaspase-3 Activation Assay-CHO cells were lysed with lysis buffer (Cell Signaling) containing a protease inhibitor mixture, as previously described (28). The extracts were centrifuged for 10 min at 14,000 ϫ g, boiled for 5 min at 100°C, and analyzed by SDS-PAGE and immunoblotting using anti-␤-tubulin antibody (Santa Cruz, CA) or antibodies for inactive caspase-3 (9665S, clone 8G10) and active caspase-3 (9661S, clone Asp 175 ).…”

Section: Methodsmentioning

confidence: 99%

Chaperone Peptides of α-Crystallin Inhibit Epithelial Cell Apoptosis, Protein Insolubilization, and Opacification in Experimental Cataracts

Nahomi

Wang

Raghavan

et al. 2013

Journal of Biological Chemistry

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Binding of Caspase-3 Prodomain to Heat Shock Protein 27 Regulates Monocyte Apoptosis by Inhibiting Caspase-3 Proteolytic Activation

Cited by 152 publications

References 36 publications

Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood–brain barrier disruption after ischemic brain injury

Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood–brain barrier disruption after ischemic brain injury

Ordering of caspases in cells undergoing apoptosis by the intrinsic pathway

Chaperone Peptides of α-Crystallin Inhibit Epithelial Cell Apoptosis, Protein Insolubilization, and Opacification in Experimental Cataracts

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