3 research outputs found
Split<sup>2</sup> Protein-Ligation Generates Active IL-6-Type Hyper-Cytokines from Inactive Precursors
Trans-signaling of
the major pro- and anti-inflammatory cytokines
Interleukin (IL)-6 and IL-11 has the unique feature to virtually activate
all cells of the body and is critically involved in chronic inflammation
and regeneration. Hyper-IL-6 and Hyper-IL-11 are single chain designer
trans-signaling cytokines, in which the cytokine and soluble receptor
units are trapped in one complex <i>via</i> a flexible peptide
linker. Albeit, Hyper-cytokines are essential tools to study trans-signaling <i>in vitro</i> and <i>in vivo</i>, the superior potency
of these designer cytokines are accompanied by undesirable stress
responses. To enable tailor-made generation of Hyper-cytokines, we
developed inactive split-cytokine-precursors adapted for posttranslational
reassembly by split-intein mediated protein trans-splicing (PTS).
We identified cutting sites within IL-6 (E<sup>134</sup>/S<sup>135</sup>) and IL-11 (G<sup>116</sup>/S<sup>117</sup>) and obtained inactive
split-Hyper-IL-6 and split-Hyper-IL-11 cytokine precursors. After
fusion with split-inteins, PTS resulted in reconstitution of active
Hyper-cytokines, which were efficiently secreted from transfected
cells. Our strategy comprises the development of a background-free
cytokine signaling system from reversibly inactivated precursor cytokines