Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs
Citation:
Clancy D.M, Henry C.M, Davidovich P.B, Sullivan G.P, Belotcerkovskaya E, Martin S.J, Production of biologically active IL-36 family cytokines through insertion of N-terminal caspase cleavage motifs, FEBS Open Bio, 6, 4, 2016, 338 - 348Download Item:
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
a
, IL-36
b
and IL-36
c
cytokines were
highly soluble and were readily processed and activated by caspase-3. Cas-
pase-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
Sponsor
Grant Number
Science Foundation Ireland (SFI)
PI (14/IA/ 2622
Author's Homepage:
http://people.tcd.ie/martinsjDescription:
PUBLISHEDExport Date: 6 January 2017
Author: MARTIN, SEAMUS
Type of material:
Journal ArticleCollections
Series/Report no:
FEBS Open Bio6
4
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Full text availableKeywords:
IL-36 cytokinesDOI:
http://dx.doi.org/10.1002/2211-5463.12044Metadata
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