Combined <i>in vitro</i> and cell‐based selection display method producing specific binders against <scp>IL</scp>‐9 receptor in high yields

Huličiak, Maroš; Biedermanová, Lada; Berdár, Daniel; Herynek, Štěpán; Kolářová, Lucie; Tomala, Jakub; Mikulecký, Pavel; Schneider, Bohdan

doi:10.1111/febs.16726

Cited by 1 publication

(1 citation statement)

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“…Overall, our results suggest the feasibility of regulating interactions involving protein partners from the interleukin family, IL-24 and IL-20R2, with UV light and photocaged tyrosines introduced at certain residue positions. In the future, other cytokine/receptor pairs and non-antibody-based protein scaffolds (Kolářová et al, 2021;Pham et al, 2021;Huličiak et al, 2023) may benefit from a similar approach and offer excellent interaction control.…”

Section: Monitoring Il-24 -Il-20r2 Interactions By Biophysical Method...mentioning

confidence: 99%

Regulation of IL-24/IL-20R2 complex formation using photocaged tyrosines and UV light

Pham

Zahradnı́k

Kolářová

et al. 2023

Front. Mol. Biosci.

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Human interleukin 24 (IL-24) is a multifunctional cytokine that represents an important target for autoimmune diseases and cancer. Since the biological functions of IL-24 depend on interactions with membrane receptors, on-demand regulation of the affinity between IL-24 and its cognate partners offers exciting possibilities in basic research and may have applications in therapy. As a proof-of-concept, we developed a strategy based on recombinant soluble protein variants and genetic code expansion technology to photocontrol the binding between IL-24 and one of its receptors, IL-20R2. Screening of non-canonical ortho-nitrobenzyl-tyrosine (NBY) residues introduced at several positions in both partners was done by a combination of biophysical and cell signaling assays. We identified one position for installing NBY, tyrosine70 of IL-20R2, which results in clear impairment of heterocomplex assembly in the dark. Irradiation with 365-nm light leads to decaging and reconstitutes the native tyrosine of the receptor that can then associate with IL-24. Photocaged IL-20R2 may be useful for the spatiotemporal control of the JAK/STAT phosphorylation cascade.

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Section: Monitoring Il-24 -Il-20r2 Interactions By Biophysical Method...mentioning

confidence: 99%

Regulation of IL-24/IL-20R2 complex formation using photocaged tyrosines and UV light

Pham

Zahradnı́k

Kolářová

et al. 2023

Front. Mol. Biosci.

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show abstract

Combined in vitro and cell‐based selection display method producing specific binders against IL‐9 receptor in high yields

Abstract: Table S1. Protein sequence of human IL-9Rα extracellular domain containing C-terminal 6xHis purification tag, and cloning sites.

Cited by 1 publication

References 33 publications

Regulation of IL-24/IL-20R2 complex formation using photocaged tyrosines and UV light

Regulation of IL-24/IL-20R2 complex formation using photocaged tyrosines and UV light

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