{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T10:24:43Z","timestamp":1784543083227,"version":"3.55.0"},"reference-count":31,"publisher":"Springer Science and Business Media LLC","issue":"7","license":[{"start":{"date-parts":[[2015,7,9]],"date-time":"2015-07-09T00:00:00Z","timestamp":1436400000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2015,7,9]],"date-time":"2015-07-09T00:00:00Z","timestamp":1436400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Cell Death Dis"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The human inflammatory caspases, including caspase-1, -4, -5 and -12, are considered as key regulators of innate immunity protecting from sepsis and numerous inflammatory diseases. Caspase-1 is activated by proximity-induced dimerization following recruitment to inflammasomes but the roles of the remaining inflammatory caspases in inflammasome assembly are unclear. Here, we use caspase bimolecular fluorescence complementation to visualize the assembly of inflammasomes and dimerization of inflammatory caspases in single cells. We observed caspase-1 dimerization induced by the coexpression of a range of inflammasome proteins and by lipospolysaccharide (LPS) treatment in primary macrophages. Caspase-4 and -5 were only dimerized by select inflammasome proteins, whereas caspase-12 dimerization was not detected by any investigated treatment. Strikingly, we determined that certain inflammasome proteins could induce heterodimerization of caspase-1 with caspase-4 or -5. Caspase-5 homodimerization and caspase-1\/-5 heterodimerization was also detected in LPS-primed primary macrophages in response to cholera toxin subunit B. The subcellular localization and organization of the inflammasome complexes varied markedly depending on the upstream trigger and on which caspase or combination of caspases were recruited. Three-dimensional imaging of the ASC (apoptosis-associated speck-like protein containing a caspase recruitment domain)\/caspase-1 complexes revealed a large spherical complex of ASC with caspase-1 dimerized on the outer surface. In contrast, NALP1 (NACHT leucine-rich repeat protein 1)\/caspase-1 complexes formed large filamentous structures. These results argue that caspase-1, -4 or -5 can be recruited to inflammasomes under specific circumstances, often leading to distinctly organized and localized complexes that may impact the functions of these proteases.<\/jats:p>","DOI":"10.1038\/cddis.2015.186","type":"journal-article","created":{"date-parts":[[2015,7,9]],"date-time":"2015-07-09T12:44:42Z","timestamp":1436445882000},"page":"e1813-e1813","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["Single-cell imaging of inflammatory caspase dimerization reveals differential recruitment to inflammasomes"],"prefix":"10.1038","volume":"6","author":[{"given":"M G","family":"Sanders","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M J","family":"Parsons","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A G A","family":"Howard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S R","family":"Fassio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J A","family":"Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L","family":"Bouchier-Hayes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2015,7,9]]},"reference":[{"key":"BFcddis2015186_CR1","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1126\/science.1373520","volume":"256","author":"DP Cerretti","year":"1992","unstructured":"Cerretti DP, Kozlosky CJ, Mosley B, Nelson N, Van Ness K, Greenstreet TA et al. Molecular cloning of the interleukin-1 beta converting enzyme. Science 1992; 256: 97\u2013100.","journal-title":"Science"},{"key":"BFcddis2015186_CR2","doi-asserted-by":"publisher","first-page":"619","DOI":"10.1038\/386619a0","volume":"386","author":"T Ghayur","year":"1997","unstructured":"Ghayur T, Banerjee S, Hugunin M, Butler D, Herzog L, Carter A et al. Caspase-1 processes IFN-gamma-inducing factor and regulates LPS-induced IFN-gamma production. Nature 1997; 386: 619\u2013623.","journal-title":"Nature"},{"key":"BFcddis2015186_CR3","doi-asserted-by":"publisher","first-page":"3502","DOI":"10.1172\/JCI40599","volume":"119","author":"A Stutz","year":"2009","unstructured":"Stutz A, Golenbock DT, Latz E . Inflammasomes: too big to miss. J Clin Invest 2009; 119: 3502\u20133511.","journal-title":"J Clin Invest"},{"key":"BFcddis2015186_CR4","doi-asserted-by":"publisher","first-page":"615","DOI":"10.1016\/j.coi.2007.09.002","volume":"19","author":"V Petrilli","year":"2007","unstructured":"Petrilli V, Dostert C, Muruve DA, Tschopp J . The inflammasome: a danger sensing complex triggering innate immunity. Curr Opin Immunol 2007; 19: 615\u2013622.","journal-title":"Curr Opin Immunol"},{"key":"BFcddis2015186_CR5","doi-asserted-by":"publisher","first-page":"1273","DOI":"10.1038\/sj.cdd.4400774","volume":"7","author":"J Bertin","year":"2000","unstructured":"Bertin J, DiStefano PS . The PYRIN domain: a novel motif found in apoptosis and inflammation proteins. Cell Death Differ 2000; 7: 1273\u20131274.","journal-title":"Cell Death Differ"},{"key":"BFcddis2015186_CR6","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1016\/S0968-0004(97)01043-8","volume":"22","author":"K Hofmann","year":"1997","unstructured":"Hofmann K, Bucher P, Tschopp J . The CARD domain: a new apoptotic signalling motif. Trends Biochem Sci 1997; 22: 155\u2013156.","journal-title":"Trends Biochem Sci"},{"key":"BFcddis2015186_CR7","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1016\/S1097-2765(02)00599-3","volume":"10","author":"F Martinon","year":"2002","unstructured":"Martinon F, Burns K, Tschopp J . The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell 2002; 10: 417\u2013426.","journal-title":"Mol Cell"},{"key":"BFcddis2015186_CR8","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/S1074-7613(04)00046-9","volume":"20","author":"L Agostini","year":"2004","unstructured":"Agostini L, Martinon F, Burns K, McDermott MF, Hawkins PN, Tschopp J . NALP3 forms an IL-1beta-processing inflammasome with increased activity in Muckle-Wells autoinflammatory disorder. Immunity 2004; 20: 319\u2013325.","journal-title":"Immunity"},{"key":"BFcddis2015186_CR9","doi-asserted-by":"publisher","first-page":"33835","DOI":"10.1074\/jbc.274.48.33835","volume":"274","author":"J Masumoto","year":"1999","unstructured":"Masumoto J, Taniguchi S, Ayukawa K, Sarvotham H, Kishino T, Niikawa N et al. ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. J Biol Chem 1999; 274: 33835\u201333838.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR10","doi-asserted-by":"publisher","first-page":"21119","DOI":"10.1074\/jbc.C200179200","volume":"277","author":"SM Srinivasula","year":"2002","unstructured":"Srinivasula SM, Poyet JL, Razmara M, Datta P, Zhang Z, Alnemri ES . The PYRIN-CARD protein ASC is an activating adaptor for caspase-1. J Biol Chem 2002; 277: 21119\u201321122.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR11","doi-asserted-by":"publisher","first-page":"28309","DOI":"10.1074\/jbc.C100250200","volume":"276","author":"JL Poyet","year":"2001","unstructured":"Poyet JL, Srinivasula SM, Tnani M, Razmara M, Fernandes-Alnemri T, Alnemri ES . Identification of Ipaf, a human caspase-1-activating protein related to Apaf-1. J Biol Chem 2001; 276: 28309\u201328313.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR12","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1038\/nature02664","volume":"430","author":"S Mariathasan","year":"2004","unstructured":"Mariathasan S, Newton K, Monack DM, Vucic D, French DM, Lee WP et al. Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf. Nature 2004; 430: 213\u2013218.","journal-title":"Nature"},{"key":"BFcddis2015186_CR13","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1038\/ni1344","volume":"7","author":"EA Miao","year":"2006","unstructured":"Miao EA, Alpuche-Aranda CM, Dors M, Clark AE, Bader MW, Miller SI et al. Cytoplasmic flagellin activates caspase-1 and secretion of interleukin 1beta via Ipaf. Nat Immunol 2006; 7: 569\u2013575.","journal-title":"Nat Immunol"},{"key":"BFcddis2015186_CR14","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1038\/sj.cdd.4402026","volume":"14","author":"AM Scott","year":"2007","unstructured":"Scott AM, Saleh M . The inflammatory caspases: guardians against infections and sepsis. Cell Death Differ 2007; 14: 23\u201331.","journal-title":"Cell Death Differ"},{"key":"BFcddis2015186_CR15","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1038\/nature02451","volume":"429","author":"M Saleh","year":"2004","unstructured":"Saleh M, Vaillancourt JP, Graham RK, Huyck M, Srinivasula SM, Alnemri ES et al. Differential modulation of endotoxin responsiveness by human caspase-12 polymorphisms. Nature 2004; 429: 75\u201379.","journal-title":"Nature"},{"key":"BFcddis2015186_CR16","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1038\/nature10558","volume":"479","author":"N Kayagaki","year":"2011","unstructured":"Kayagaki N, Warming S, Lamkanfi M, Vande Walle L, Louie S, Dong J et al. Non-canonical inflammasome activation targets caspase-11. Nature 2011; 479: 117\u2013121.","journal-title":"Nature"},{"key":"BFcddis2015186_CR17","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1038\/nature13683","volume":"514","author":"J Shi","year":"2014","unstructured":"Shi J, Zhao Y, Wang Y, Gao W, Ding J, Li P et al. Inflammatory caspases are innate immune receptors for intracellular LPS. Nature 2014; 514: 187\u2013192.","journal-title":"Nature"},{"key":"BFcddis2015186_CR18","doi-asserted-by":"publisher","first-page":"21777","DOI":"10.1074\/jbc.R800084200","volume":"284","author":"C Pop","year":"2009","unstructured":"Pop C, Salvesen GS . Human caspases: activation, specificity, and regulation. J Biol Chem 2009; 284: 21777\u201321781.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR19","doi-asserted-by":"publisher","first-page":"830","DOI":"10.1016\/j.molcel.2009.07.023","volume":"35","author":"L Bouchier-Hayes","year":"2009","unstructured":"Bouchier-Hayes L, Oberst A, McStay GP, Connell S, Tait SW, Dillon CP et al. Characterization of cytoplasmic caspase-2 activation by induced proximity. Mol Cell 2009; 35: 830\u2013840.","journal-title":"Mol Cell"},{"key":"BFcddis2015186_CR20","doi-asserted-by":"publisher","first-page":"39920","DOI":"10.1074\/jbc.M007255200","volume":"275","author":"XY Lin","year":"2000","unstructured":"Lin XY, Choi MS, Porter AG . Expression analysis of the human caspase-1 subfamily reveals specific regulation of the CASP5 gene by lipopolysaccharide and interferon-gamma. J Biol Chem 2000; 275: 39920\u201339926.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR21","doi-asserted-by":"publisher","first-page":"613","DOI":"10.1042\/BJ20121198","volume":"449","author":"M Proell","year":"2013","unstructured":"Proell M, Gerlic M, Mace PD, Reed JC, Riedl SJ . The CARD plays a critical role in ASC foci formation and inflammasome signalling. Biochem J 2013; 449: 613\u2013621.","journal-title":"Biochem J"},{"key":"BFcddis2015186_CR22","doi-asserted-by":"publisher","first-page":"1590","DOI":"10.1038\/sj.cdd.4402194","volume":"14","author":"T Fernandes-Alnemri","year":"2007","unstructured":"Fernandes-Alnemri T, Wu J, Yu JW, Datta P, Miller B, Jankowski W et al. The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation. Cell Death Differ 2007; 14: 1590\u20131604.","journal-title":"Cell Death Differ"},{"key":"BFcddis2015186_CR23","doi-asserted-by":"publisher","first-page":"17907","DOI":"10.1074\/jbc.272.29.17907","volume":"272","author":"NA Thornberry","year":"1997","unstructured":"Thornberry NA, Rano TA, Peterson EP, Rasper DM, Timkey T, Garcia-Calvo M et al. A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J Biol Chem 1997; 272: 17907\u201317911.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR24","doi-asserted-by":"publisher","first-page":"15870","DOI":"10.1074\/jbc.270.26.15870","volume":"270","author":"NA Munday","year":"1995","unstructured":"Munday NA, Vaillancourt JP, Ali A, Casano FJ, Miller DK, Molineaux SM et al. Molecular cloning and pro-apoptotic activity of ICErelII and ICErelIII, members of the ICE\/CED-3 family of cysteine proteases. J Biol Chem 1995; 270: 15870\u201315876.","journal-title":"J Biol Chem"},{"key":"BFcddis2015186_CR25","doi-asserted-by":"publisher","first-page":"880","DOI":"10.1074\/mcp.M900528-MCP200","volume":"9","author":"NJ Agard","year":"2010","unstructured":"Agard NJ, Maltby D, Wells JA . Inflammatory stimuli regulate caspase substrate profiles. Mol Cell Proteom 2010; 9: 880\u2013893.","journal-title":"Mol Cell Proteom"},{"key":"BFcddis2015186_CR26","doi-asserted-by":"publisher","first-page":"335","DOI":"10.4049\/jimmunol.1303424","volume":"193","author":"Y Kajiwara","year":"2014","unstructured":"Kajiwara Y, Schiff T, Voloudakis G, Gama Sosa MA, Elder G, Bozdagi O et al. A critical role for human caspase-4 in endotoxin sensitivity. J Immunol 2014; 193: 335\u2013343.","journal-title":"J Immunol"},{"key":"BFcddis2015186_CR27","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1042\/BJ20101738","volume":"433","author":"C Pop","year":"2011","unstructured":"Pop C, Oberst A, Drag M, Van Raam BJ, Riedl SJ, Green DR et al. FLIP(L) induces caspase 8 activity in the absence of interdomain caspase 8 cleavage and alters substrate specificity. Biochem J 2011; 433: 447\u2013457.","journal-title":"Biochem J"},{"key":"BFcddis2015186_CR28","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1006\/bbrc.2000.4190","volume":"280","author":"J Masumoto","year":"2001","unstructured":"Masumoto J, Taniguchi S, Sagara J . Pyrin N-terminal homology domain- and caspase recruitment domain-dependent oligomerization of ASC. Biochem Biophys Res Commun 2001; 280: 652\u2013655.","journal-title":"Biochem Biophys Res Commun"},{"key":"BFcddis2015186_CR29","doi-asserted-by":"publisher","first-page":"1193","DOI":"10.1016\/j.cell.2014.02.008","volume":"156","author":"A Lu","year":"2014","unstructured":"Lu A, Magupalli VG, Ruan J, Yin Q, Atianand MK, Vos MR et al. Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. Cell 2014; 156: 1193\u20131206.","journal-title":"Cell"},{"key":"BFcddis2015186_CR30","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1016\/j.cell.2004.05.004","volume":"117","author":"F Martinon","year":"2004","unstructured":"Martinon F, Tschopp J . Inflammatory caspases: linking an intracellular innate immune system to autoinflammatory diseases. Cell 2004; 117: 561\u2013574.","journal-title":"Cell"},{"key":"BFcddis2015186_CR31","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1038\/ng756","volume":"29","author":"HM Hoffman","year":"2001","unstructured":"Hoffman HM, Mueller JL, Broide DH, Wanderer AA, Kolodner RD . Mutation of a new gene encoding a putative pyrin-like protein causes familial cold autoinflammatory syndrome and Muckle\u2013Wells syndrome. Nat Genet 2001; 29: 301\u2013305.","journal-title":"Nat Genet"}],"container-title":["Cell Death &amp; Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/cddis2015186.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/cddis2015186","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/cddis2015186.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T10:06:15Z","timestamp":1672913175000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/cddis2015186"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,7,9]]},"references-count":31,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2015,7]]}},"alternative-id":["BFcddis2015186"],"URL":"https:\/\/doi.org\/10.1038\/cddis.2015.186","relation":{},"ISSN":["2041-4889"],"issn-type":[{"value":"2041-4889","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,7,9]]},"assertion":[{"value":"11 May 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 May 2015","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 July 2015","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no conflict of interest.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}