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Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs

(2016) FEBS OPEN BIO. 6(4). p.338-348
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Abstract
Recent evidence has strongly implicated IL-36 cytokines as key initiators of inflammation in the skin barrier. IL-36 cytokines belong to the extended IL-1 family and, similar to most members of this family, are expressed as inactive precursors that require proteolytic processing for activation. Because the proteases responsible for activation of members of the IL-36 subfamily have not been reported, we have developed a method for the production of biologically active IL-36 through introduction of a caspase cleavage motif, DEVD, within the N-termini of these cytokines. Here, we show that DEVD-modified IL-36 alpha, IL-36 beta and IL-36 gamma cytokines were highly soluble and were readily processed and activated by caspase-3. Caspase-3-processed IL-36 family cytokines exhibited robust biological activity on a range of responsive cell types, including primary keratinocytes. We also generated specific polyclonal antibodies against all three IL-36 family members through immunization with purified recombinant IL-36 cytokines. The modified forms of IL-36 described herein will be useful for production of large quantities of biologically active IL-36 for structure and function studies on these important proinflammatory cytokines.
Keywords
caspase, cell death, IL-1 family, IL-36, inflammation, protease, GENERALIZED PUSTULAR PSORIASIS, INTERLEUKIN-1 FAMILY, T-CELLS, KAPPA-B, PROTEASE, ANTAGONIST, LIGANDS, PURIFICATION, ACTIVATION, EXPRESSION

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MLA
Clancy, Danielle, et al. “Production of Biologically Active IL-36 Family Cytokines through Insertion of N-Terminal Caspase Cleavage Motifs.” FEBS OPEN BIO, vol. 6, no. 4, 2016, pp. 338–48, doi:10.1002/2211-5463.12044.
APA
Clancy, D., Henry, C. M., Davidovich, P. B., Sullivan, G. P., Belotcerkovskaya, E., & Martin, S. J. (2016). Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs. FEBS OPEN BIO, 6(4), 338–348. https://doi.org/10.1002/2211-5463.12044
Chicago author-date
Clancy, Danielle, Conor M. Henry, Pavel B. Davidovich, Graeme P. Sullivan, Ekaterina Belotcerkovskaya, and Seamus J. Martin. 2016. “Production of Biologically Active IL-36 Family Cytokines through Insertion of N-Terminal Caspase Cleavage Motifs.” FEBS OPEN BIO 6 (4): 338–48. https://doi.org/10.1002/2211-5463.12044.
Chicago author-date (all authors)
Clancy, Danielle, Conor M. Henry, Pavel B. Davidovich, Graeme P. Sullivan, Ekaterina Belotcerkovskaya, and Seamus J. Martin. 2016. “Production of Biologically Active IL-36 Family Cytokines through Insertion of N-Terminal Caspase Cleavage Motifs.” FEBS OPEN BIO 6 (4): 338–348. doi:10.1002/2211-5463.12044.
Vancouver
1.
Clancy D, Henry CM, Davidovich PB, Sullivan GP, Belotcerkovskaya E, Martin SJ. Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs. FEBS OPEN BIO. 2016;6(4):338–48.
IEEE
[1]
D. Clancy, C. M. Henry, P. B. Davidovich, G. P. Sullivan, E. Belotcerkovskaya, and S. J. Martin, “Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs,” FEBS OPEN BIO, vol. 6, no. 4, pp. 338–348, 2016.
@article{8727874,
  abstract     = {{Recent evidence has strongly implicated IL-36 cytokines as key initiators of inflammation in the skin barrier. IL-36 cytokines belong to the extended IL-1 family and, similar to most members of this family, are expressed as inactive precursors that require proteolytic processing for activation. Because the proteases responsible for activation of members of the IL-36 subfamily have not been reported, we have developed a method for the production of biologically active IL-36 through introduction of a caspase cleavage motif, DEVD, within the N-termini of these cytokines. Here, we show that DEVD-modified IL-36 alpha, IL-36 beta and IL-36 gamma cytokines were highly soluble and were readily processed and activated by caspase-3. Caspase-3-processed IL-36 family cytokines exhibited robust biological activity on a range of responsive cell types, including primary keratinocytes. We also generated specific polyclonal antibodies against all three IL-36 family members through immunization with purified recombinant IL-36 cytokines. The modified forms of IL-36 described herein will be useful for production of large quantities of biologically active IL-36 for structure and function studies on these important proinflammatory cytokines.}},
  author       = {{Clancy, Danielle and Henry, Conor M. and Davidovich, Pavel B. and Sullivan, Graeme P. and Belotcerkovskaya, Ekaterina and Martin, Seamus J.}},
  issn         = {{2211-5463}},
  journal      = {{FEBS OPEN BIO}},
  keywords     = {{caspase,cell death,IL-1 family,IL-36,inflammation,protease,GENERALIZED PUSTULAR PSORIASIS,INTERLEUKIN-1 FAMILY,T-CELLS,KAPPA-B,PROTEASE,ANTAGONIST,LIGANDS,PURIFICATION,ACTIVATION,EXPRESSION}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{338--348}},
  title        = {{Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs}},
  url          = {{http://dx.doi.org/10.1002/2211-5463.12044}},
  volume       = {{6}},
  year         = {{2016}},
}

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