The Caspase-1 Digestome Identifies the Glycolysis Pathway as a Target during Infection and Septic Shock

Caspase-1 is an essential effector of inflammation, pyroptosis, and septic shock. Few caspase-1 substrates have been identified to date, and these substrates do not account for its wide range of actions. To understand the function of caspase-1, we initiated the systematic identification of its cellular substrates. Using the diagonal gel proteomic approach, we identified 41 proteins that are directly cleaved by caspase-1. Among these were chaperones, cytoskeletal and translation machinery proteins, and proteins involved in immunity. A series of unexpected proteins along the glycolysis pathway were also identified, including aldolase, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, α-enolase, and pyruvate kinase. With the exception of the latter, the identified glycolysis enzymes were specifically cleaved in vitro by recombinant caspase-1, but not caspase-3. The enzymatic activity of wild-type glyceraldehyde-3-phosphate dehydrogenase, but not a non-cleavable mutant, was dampened by caspase-1 processing. In vivo, stimuli that fully activated caspase-1, including Salmonella typhimurium infection and septic shock, caused a pronounced processing of these proteins in the macrophage and diaphragm muscle, respectively. Notably, these stimuli inhibited glycolysis in wild-type cells compared with caspase-1-deficient cells. The systematic characterization of caspase-1 substrates identifies the glycolysis pathway as a caspase-1 target and provides new insights into its function during pyroptosis and septic shock. Caspase-1 is an essential effector of inflammation, pyroptosis, and septic shock. Few caspase-1 substrates have been identified to date, and these substrates do not account for its wide range of actions. To understand the function of caspase-1, we initiated the systematic identification of its cellular substrates. Using the diagonal gel proteomic approach, we identified 41 proteins that are directly cleaved by caspase-1. Among these were chaperones, cytoskeletal and translation machinery proteins, and proteins involved in immunity. A series of unexpected proteins along the glycolysis pathway were also identified, including aldolase, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, α-enolase, and pyruvate kinase. With the exception of the latter, the identified glycolysis enzymes were specifically cleaved in vitro by recombinant caspase-1, but not caspase-3. The enzymatic activity of wild-type glyceraldehyde-3-phosphate dehydrogenase, but not a non-cleavable mutant, was dampened by caspase-1 processing. In vivo, stimuli that fully activated caspase-1, including Salmonella typhimurium infection and septic shock, caused a pronounced processing of these proteins in the macrophage and diaphragm muscle, respectively. Notably, these stimuli inhibited glycolysis in wild-type cells compared with caspase-1-deficient cells. The systematic characterization of caspase-1 substrates identifies the glycolysis pathway as a caspase-1 target and provides new insights into its function during pyroptosis and septic shock. Caspases are aspartate-specific cysteine proteases known for their function in regulating programmed cell death and inflammation. Phylogenetically, they are subdivided into the CED3-related enzymes that initiate and execute cell death and the caspase-1-related proteins that process and mature cytokines, viz. IL-1β, 2The abbreviations used are:ILinterleukinTIMtriose-phosphate isomeraseGAPDHglyceraldehyde-3-phosphate dehydrogenaseCHAPS3-(3-cholamidopropyl) dimethylammonio-1-propanesulfonic acidLPSlipopolysaccharideBisTris2-bis(2-hydroxyethyl) amino-2-(hydroxymethyl) propane-1,3-diolTBSTris-buffered salineMALDI-TOFmatrix-assisted laser desorption ionization time-of-flight. IL-18, and IL-33. Apoptotic caspases execute cell death through the restricted cleavage of key cellular proteins required to maintain cell viability, resulting in the morphological changes observed during apoptosis, including membrane blebbing, nuclear condensation, and cytoskeletal dismantling (1Nicholson D.W. Cell Death Differ. 1999; 6: 1028-1042Crossref PubMed Scopus (1306) Google Scholar). interleukin triose-phosphate isomerase glyceraldehyde-3-phosphate dehydrogenase 3-(3-cholamidopropyl) dimethylammonio-1-propanesulfonic acid lipopolysaccharide 2-bis(2-hydroxyethyl) amino-2-(hydroxymethyl) propane-1,3-diol Tris-buffered saline matrix-assisted laser desorption ionization time-of-flight Caspase-1 is essential during inflammation because of its role in the activation of cytokine signaling pathways. With the exception of its cytokine substrates, very little is known regarding the spectrum of cellular proteins it targets upon full activation. Similar to other caspases, caspase-1 is found in cells as an inactive precursor and is activated in response to inflammatory triggers, including pathogen-derived molecules, as well as danger signals released from infected or dying cells (2Scott A.M. Saleh M. Cell Death Differ. 2007; 14: 23-31Crossref PubMed Scopus (105) Google Scholar). Caspase-1 activation is achieved in a macromolecular complex known as the inflammasome through its recruitment to a scaffolding molecule generally via the adaptor ASC (3Mariathasan S. Monack D.M. Nat. Rev. Immunol. 2007; 7: 31-40Crossref PubMed Scopus (701) Google Scholar). Scaffolding molecules that activate caspase-1 within the inflammasome belong to the cytosolic Nod-like family of pathogen recognition receptors and include Nalp1–3, Ipaf, and Naip5 (4Tschopp J. Martinon F. Burns K. Nat. Rev. Mol. Cell Biol. 2003; 4: 95-104Crossref PubMed Scopus (601) Google Scholar). More recently, a distinct caspase-1 activation platform, the ASC pyroptosome, has been characterized (5Fernandes-Alnemri T. Wu J. Yu J.W. Datta P. Miller B. Jankowski W. Rosenberg S. Zhang J. Alnemri E.S. Cell Death Differ. 2007; 14: 1590-1604Crossref PubMed Scopus (740) Google Scholar). It differs from the inflammasome in that it does not contain a Nod-like scaffolding protein but is assembled through the oligomerization of ASC. Although caspase-1 is activated transiently within the Nalp inflammasomes, resulting in controlled inflammation and the restricted processing of cytokine substrates, it is fully activated within the pyroptosome, leading to a distinct form of inflammatory cell death known as pyroptosis (6Saleh M. Green D.R. Cell Death Differ. 2007; 14: 1559-1560Crossref PubMed Scopus (15) Google Scholar). Pyroptosis exhibits features common to both apoptosis and necrosis, including nuclear condensation, loss of mitochondrial membrane potential, and membrane swelling (5Fernandes-Alnemri T. Wu J. Yu J.W. Datta P. Miller B. Jankowski W. Rosenberg S. Zhang J. Alnemri E.S. Cell Death Differ. 2007; 14: 1590-1604Crossref PubMed Scopus (740) Google Scholar). It was initially observed in macrophages infected with the intracellular pathogen Salmonella typhimurium (7Monack D.M. Navarre W.W. Falkow S. Microbes Infect. 2001; 3: 1201-1212Crossref PubMed Scopus (99) Google Scholar) but was later found to occur in response to infection with other intracellular pathogens (8Fink S.L. Cookson B.T. Infect. Immun. 2005; 73: 1907-1916Crossref PubMed Scopus (1518) Google Scholar, 9Swanson M.S. Molofsky A.B. Autophagy. 2005; 1: 174-176Crossref PubMed Scopus (47) Google Scholar). Because only a few substrates for caspase-1 have been identified thus far, the mechanism by which caspase-1 kills the cell via pyroptosis remains obscure. Another instance in which caspase-1 is activated is during septic shock (10Kuida K. Lippke J.A. Ku G. Harding M.W. Livingston D.J. Su M.S. Flavell R.A. Science. 1995; 267: 2000-2003Crossref PubMed Scopus (1461) Google Scholar, 11Li P. Allen H. Banerjee S. Franklin S. Herzog L. Johnston C. McDowell J. Paskind M. Rodman L. Salfeld J. Towne E. Tracey D. Wardwell S. Wei F.-Y. Wong W. Kamen R. Seshadri T. Cell. 1995; 80: 401-411Abstract Full Text PDF PubMed Scopus (1314) Google Scholar). Hyperproduction of cytokines alone does not account for the totality of caspase-1 effects during septic shock. One characteristic of this condition is an impaired ventilatory muscle contractility that causes ventilatory failure and respiratory arrest (12Hussain S.N. Mol. Cell. Biochem. 1998; 179: 125-134Crossref PubMed Scopus (87) Google Scholar), and caspase-1-deficient mice, but not IL-1β/IL-18 double knock-out mice, are protected from this condition (13Sarkar A. Hall M.W. Exline M. Hart J. Knatz N. Gatson N.T. Wewers M.D. Am. J. Respir. Crit. Care Med. 2006; 174: 1003-1010Crossref PubMed Scopus (122) Google Scholar). To understand the role of caspase-1 in pyroptosis and septic shock, we sought to identify its cellular substrates. Here, we used the diagonal gel proteomic approach and identified multiple caspase-1 direct targets. These included structural proteins, chaperones, translation machinery proteins, and proteins involved in immunity. A series of unexpected proteins along the glycolysis pathway were also identified as caspase-1 substrates, including aldolase, TIM, GAPDH, enolase, and pyruvate kinase. Here, we show that Salmonella infection, which fully activates caspase-1 and induces pyroptosis (7Monack D.M. Navarre W.W. Falkow S. Microbes Infect. 2001; 3: 1201-1212Crossref PubMed Scopus (99) Google Scholar), caused a pronounced degradation of these glycolysis enzymes and lowered the glycolytic rate of wild-type macrophages, but not caspase-1-deficient cells. Similarly, we show that the glycolysis enzymes were processed in the diaphragm muscle of wild-type septic mice. Detailed analysis of the caspase-1 substrates is likely to shed light on caspase-1 function in pyroptotic cell death and septic shock. Antibodies and Reagents—Anti-α-enolase (1:2000 dilution in muscle and 1:1000 dilution in monocytes; catalog no. sc-7455), anti-aldolase (1:2000 dilution in muscle and 1:1000 dilution in monocytes; catalog no. sc-12059), anti-TIM (1:1000 dilution; catalog no. sc-22031), and anti-caspase-1 p10 subunit (1:500 dilution in muscle and 1:1000 dilution in monocytes; catalog no. sc-514) antibodies were purchased from Santa Cruz Biotechnology, Inc. Anti-GAPDH antibody (1:1000 dilution; Ab9485) was obtained from Abcam. Anti-IL-1β antibody (1:1000 dilution; catalog no. 2022) was purchased from Cell Signaling Technology. Human recombinant caspase-1 was from Merck. TRIzol RNA extraction reagent, oligo(dT)12–18 primers, random hexamers, and Moloney murine leukemia virus reverse transcriptase were from Invitrogen. RNase inhibitor and an in vitro transcription/translation kit were from Promega Corp. The QuikChange site-directed mutagenesis kit was from Stratagene. Diagonal Gel—18 × 106 THP-1 cells or human peripheral blood mononuclear cells were lysed in 200 μl of Laemmli SDS loading buffer (50 mm Tris (pH 6.8), 2% SDS, 0.1% bromphenol blue, 10% glycerol, and 2.5% β-mercaptoethanol) and for the to of protein was by 10% the the protein was and in and 10% acid for in for and in for The was the gel to in buffer mm (pH mm and 10% with mm with or of recombinant caspase-1 or and in a The was with to the and in Laemmli SDS loading buffer for in a in a the was a 10% gel and the gel was in 10% acid and and proteins, which were the were from the gel and identified by the and by RNA and RNA was from THP-1 cells TRIzol × 106 cells were lysed in 200 μl of TRIzol and with μl of were via reverse or random and aldolase, TIM, and pyruvate were by from THP-1 the The were into the The from were by The was purchased from no. and into the cleavage were by site-directed mutagenesis the QuikChange site-directed mutagenesis kit the The for were and for were as GAPDH, and α-enolase, and aldolase, and α-enolase, and aldolase, and TIM, and and pyruvate and In substrates were obtained by in vitro transcription/translation the Promega or the of the was with or acid and for of the in vitro and substrates was in a in vitro and substrates by for in the or of human recombinant caspase-1 in buffer mm The cleavage was by the of Laemmli SDS loading buffer and by The gel was in 10% acid and for the was by the gel with for The gel was on a for and and the was by Salmonella for was in or on were on To for infection of THP-1 was with a and in of with The infection, were in and to to and were by for with an of saline and with and for in cell in a in a with the The were used for infection of cells. Cell and cells were a of × 106 to infection, cells × 106 were and in in a THP-1 cells were with to the cells into cells. The the and cells were and with The cells were with The cells were infected with Salmonella a of infection of or cells. were × for and for to to The was with and for an of and caspase-1 knock-out the were with of and macrophages were were with of a and macrophages were with a and an The cell was through a and in were in a of were with from E. were with The were or by of The diaphragm muscle was in of buffer mm (pH mm mm mm and mm and × for and the was for macrophages from wild-type or caspase-1 knock-out were in a of × The cells were with (50 and infected with Salmonella a of infection of of infection, the was for a kit from no. the μl of cell was with of for and the was The was an from a mm buffer (pH mm mm no. mm and from THP-1 cell in buffer of or in vitro and and The was initiated by the of to the and the activity was by was during for the from cell THP-1 cells were lysed in and proteins was in μl of buffer and with or of caspase-1 for the in vitro and or its in vitro and were in a of buffer and with or caspase-1 for × 106 cells were lysed in μl of for and for μl was a gel for and membrane for The membrane was in in 0.1% for The antibodies were as in in 0.1% and with the membrane with The was for with 0.1% The antibodies were as in in 0.1% and with the membrane for The was for with and for with 0.1% The was by the with no. for The Caspase-1 identify caspase-1 substrates, we used the diagonal gel proteomic approach C. P. B. N. Green D.R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). In this cellular proteins are on a gel which is to SDS, and in a activation buffer the of the processing of the substrates, the gel is on a resulting in the of the of cellular proteins along a diagonal and of the cleaved proteins the diagonal A and The cleavage were identified by To the we initially for targets and identified multiple known substrates along with substrates not the caspase-1 we to with cell from THP-1 macrophages as a of cellular proteins on the diagonal To for the caspase-1 we a on the caspase-1 diagonal gel and the of mature the diagonal as that the diagonal on the caspase-1 gel were and processed for with of the of the cleaved we obtained 41 proteins that were processed by caspase-1. These included proteins involved in essential for cell as cytoskeletal and and These were also by in the diagonal gel and are by caspases in during on common cellular caspase-1 and not the proteins along these pathways. of the caspase-1 substrates were also processed by the were to of the known substrates and glycolysis substrates that were identified in the caspase-1 or diagonal gel by C. J. E. D. W. R. J. Biol. Full Text Full Text PDF PubMed Scopus Google protein subunit B. A. M. S. J. Biol. Full Text Full Text PDF PubMed Scopus Google J.A. J. Biol. Full Text Full Text PDF PubMed Scopus Google S. T. C. A. J. K. M.W. K. D.J. Science. PubMed Scopus Google S. T. C. A. J. K. M.W. K. D.J. Science. PubMed Scopus Google F. R. M. Green Cell Death Differ. 2001; PubMed Scopus Google T. M. K. Miller D.W. T. 14: PubMed Scopus Google C. J. E. D. W. R. J. Biol. Full Text Full Text PDF PubMed Scopus Google T. M. K. Miller D.W. T. 14: PubMed Scopus Google S. G. A. J. 1998; Google E. A. S. A. B. B. K. A. J. Biol. 1998; Full Text Full Text PDF PubMed Scopus Google P. L. J. K. A. J. K. Nat. 2005; PubMed Scopus Google B. C. F. T. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google P. L. J. K. A. J. K. Nat. 2005; PubMed Scopus Google of cytokines and in a new Caspase-1 the proteins involved in along the glycolysis pathway were identified in the caspase-1 diagonal gel and To this we in vitro cleavage Although both caspase-1 and processed an in vitro and form of the target C. R.A. Livingston D.J. Su M.S. J. Biol. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar), only caspase-1, but not cleaved the glycolysis enzymes aldolase, TIM, GAPDH, and the other pyruvate was cleaved by both caspases Because the cleavage of by caspase-1 was that of the other glycolysis enzymes in we characterized this cleavage The of of caspase-1 to processing in a In with the full processing of by caspase-1 that required of caspase-1 cleavage of the required of the that in vivo, processing occur in which caspase-1 is fully leading to pyroptotic cell in response to stimuli that activate caspase-1, transiently leading to cytokine processing to a or for the To the of caspase-1 on we enzymatic activity in vitro in the or of caspase-1. that of cell with caspase-1 activity by to compared with the activity in the of caspase-1 a of caspase-1 on sought to identify the caspase-1 cleavage in the of the of the cleavage we cleavage and their acid to in vitro cleavage and identified the as the caspase-1 cleavage in The the processing of by caspase-1 of protein from that the caspase-1 cleavage in and the it have been To the of the caspase-1 we compared the enzymatic activity of wild-type with that of the mutant, which is to caspase-1 in the or of caspase-1. enzymatic activity that the activity of in vitro and wild-type was by in the of caspase-1, that of the non-cleavable was by only 10% compared with the activity in the of caspase-1 of cleavage by caspase-1. the of caspase-1 in processing of activity was by the caspase-1 THP-1 were with or caspase-1 for activity was by as are the from the analysis of the cleavage in that in a cleavage is to caspase-1 is the of the the caspase-1 cleavage in from the activity of the non-cleavable is in the of caspase-1. and were by in vitro transcription/translation and were used in the enzymatic as not Caspase-1 during Salmonella to the of the and of the that the glycolysis enzymes identified in the caspase-1 diagonal gel are caspase-1 substrates, the was to they are processed in that to caspase-1 activation. to this in macrophages because they are known to on glycolysis for T. R. N. R. M. N. Cell. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar) and to of their through glycolysis T. K. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, J.A. J. Biol. 1999; PubMed Scopus Google Scholar). Although glycolytic both cellular and effector of macrophages and of mitochondrial have little on these N. T. J. PubMed Scopus Google Scholar, J. PubMed Scopus Google Scholar). In an in glycolysis in activated macrophages and is required for effector including and L. J. PubMed Scopus Google Scholar, D.J. PubMed Google Scholar, D.J. 6: PubMed Scopus Google Scholar). To activate caspase-1 in vivo, we THP-1 macrophages with and or with and stimuli that resulting in caspase-1 activation within the inflammasome S. K. J. K. M. Monack D.M. 2006; PubMed Scopus Google Scholar). the other we infected THP-1 cells with the intracellular pathogen S. which activates caspase-1 within the inflammasome M. M.W. Miller A. Nat. Immunol. 2006; 7: PubMed Scopus Google Scholar, L. A. M. N. R. N. P. J. A. G. Nat. Immunol. 2006; 7: PubMed Scopus Google Scholar). In both was processed into its mature form by caspase-1 cells were with and or with and we not cleavage observed and but these were also found cells were with a condition which caspase-1 was not activated as by the of processing the other was fully processed and during Salmonella infection in a of Similarly, was cleaved and during Salmonella infection that the on the to enolase, as it was also in the were to the processing of in cells because antibodies with upon not To the degradation of the glycolysis enzymes was by caspase-1, we processing in macrophages from wild-type or caspase-1-deficient (10Kuida K. Lippke J.A. Ku G. Harding M.W. Livingston D.J. Su M.S. Flavell R.A. Science. 1995; 267: 2000-2003Crossref PubMed Scopus (1461) Google Scholar). observed that was in wild-type macrophages infected with but not in infected caspase-1-deficient macrophages the role of caspase-1 in processing in the processing of the glycolysis enzymes an on the glycolytic rate of the One of glycolytic function and inflammatory activation of macrophages is the in L. A. 1999; Full Text PDF PubMed Scopus Google Scholar). in wild-type and caspase-1-deficient cells with or or infected with that activation of caspase-1 in of as were in macrophages compared with cells in response to Salmonella infection The Caspase-1 in the in the of in that caspase-1 targets the glycolysis pathway both in vitro and in vivo, resulting in of the cellular glycolytic It has been that septic shock have in diaphragm muscle which are with a in glycolytic rate (12Hussain S.N. Mol. Cell. Biochem. 1998; 179: 125-134Crossref PubMed Scopus (87) Google Scholar). are to septic (10Kuida K. Lippke J.A. Ku G. Harding M.W. Livingston D.J. Su M.S. Flavell R.A. Science. 1995; 267: 2000-2003Crossref PubMed Scopus (1461) Google Scholar, 11Li P. Allen H. Banerjee S. Franklin S. Herzog L. Johnston C. McDowell J. Paskind M. Rodman L. Salfeld J. Towne E. Tracey D. Wardwell S. Wei F.-Y. Wong W. Kamen R. Seshadri T. Cell. 1995; 80: 401-411Abstract Full Text PDF PubMed Scopus (1314) Google Scholar), IL-1β/IL-18 double knock-out are not (13Sarkar A. Hall M.W. Exline M. Hart J. Knatz N. Gatson N.T. Wewers M.D. Am. J. Respir. Crit. Care Med. 2006; 174: 1003-1010Crossref PubMed Scopus (122) Google Scholar). that the effects of caspase-1 in septic shock are not by its role in processing these caspase-1 other substrates that to failure and the glycolysis substrates were processed by caspase-1 in the diaphragm muscle of shock. Using we the activation of caspase-1 in the diaphragm muscle in wild-type mice. the of GAPDH, aldolase, enolase, and were in the septic shock muscle in the muscle was processed into a cleavage to the caspase-1 cleavage observed in the in vitro cleavage thus that caspase-1 targets the glycolysis enzymes in the diaphragm muscle of in septic shock. In this we identified 41 cellular proteins that were processed by caspase-1 in vitro the diagonal gel proteomic Among these were glycolysis viz. aldolase, TIM, GAPDH, α-enolase, and pyruvate kinase. that these substrates were processed by caspase-1 in in vitro cleavage the caspase-1 cleavage in and that wild-type its enzymatic activity with caspase-1, a non-cleavable was to processing and its activity in the of caspase-1. we the processing of these substrates in in which caspase-1 was fully as in macrophage infection with Salmonella and in a of shock. These are with pyroptosis, a form of cell death by caspase-1. an caspase-1 and substrates are in with the that pyroptosis morphological features with apoptosis (5Fernandes-Alnemri T. Wu J. Yu J.W. Datta P. Miller B. Jankowski W. Rosenberg S. Zhang J. Alnemri E.S. Cell Death Differ. 2007; 14: 1590-1604Crossref PubMed Scopus (740) Google Scholar). also that caspase-1 as an as well as an during pyroptosis, directly cellular substrates. Because glycolysis is essential for macrophage and activation T. R. N. R. M. N. Cell. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar), the cleavage of the glycolysis substrates, which in of cellular is to an essential cell proteins are by caspase-1 in this it was not to with non-cleavable to they glycolysis and cell in the diaphragm muscle show that these glycolysis enzymes are cleaved by caspase-1. that the processing of these proteins to the loss of muscle contractility that in septic shock. It is to that during Salmonella infection, was fully and that the degradation not occur in caspase-1-deficient cells It is that the caspase-1 cleavage were for has been to occur for other substrates S. C. S. M. M. M. C. R. Cell Death Differ. 2006; PubMed Scopus Google Scholar) and a mechanism by which the cell In to the diagonal gel and in C. P. B. N. Green D.R. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, multiple have been used to identify substrates. These the of an recombinant to an in vitro and H. H. J. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar), that cell from or dying cells C. J. E. D. W. R. J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar), and of proteins in cells apoptosis to P. L. J. K. A. J. K. Nat. 2005; PubMed Scopus Google Scholar). of proteins by transcription/translation has in that the and for substrates. is that proteins and not and the of protein are in that caspases are activated other proteases also leading to protein processing. this was observed in the approach, in protein processing during apoptosis P. L. J. K. A. J. K. Nat. 2005; PubMed Scopus Google Scholar). The of the diagonal gel approach is the of cell of in vitro and proteins and of which in direct processing of targets. Because the used in the the proteins in the we caspase-1 substrates to in the These substrates as well by other as the of cells and stimuli the and of the substrates, cellular and the rate which these substrates are In the diagonal gel proteomic approach, we 41 caspase-1 substrates. that the glycolysis pathway is specifically by caspase-1 in which it is fully leading to pyroptosis as in macrophages infected with Salmonella as well as in the diaphragm muscle during shock. It is to that was found to a protein required for the of during cell death A. S. S. L. P. A. B. P. Green D.R. Cell. 2007; Full Text Full Text PDF PubMed Scopus Google Scholar). It is to that caspase-1 targets not only to glycolysis but also to its function in which is with cell In this of the cellular proteins cleaved by caspase-1, its cytokine substrates, and to the mechanism by which caspase-1 with