@misc{10481/93227,
year = {2024},
month = {5},
url = {https://hdl.handle.net/10481/93227},
abstract = {Plakophilin 1 (PKP1), a member of the p120ctn subfamily of the armadillo (ARM)-repeatcontaining
proteins, is an important structural component of cell–cell adhesion scaffolds although it
can also be ubiquitously found in the cytoplasm and the nucleus. RYBP (RING 1A and YY1 binding
protein) is a multifunctional intrinsically disordered protein (IDP) best described as a transcriptional
regulator. Both proteins are involved in the development and metastasis of several types of tumors.
We studied the binding of the armadillo domain of PKP1 (ARM-PKP1) with RYBP by using in
cellulo methods, namely immunofluorescence (IF) and proximity ligation assay (PLA), and in vitro
biophysical techniques, namely fluorescence, far-ultraviolet (far-UV) circular dichroism (CD), and
isothermal titration calorimetry (ITC). We also characterized the binding of the two proteins by using
in silico experiments. Our results showed that there was binding in tumor and non-tumoral cell lines.
Binding in vitro between the two proteins was also monitored and found to occur with a dissociation
constant in the low micromolar range (~10 μM). Finally, in silico experiments provided additional
information on the possible structure of the binding complex, especially on the binding ARM-PKP1
hot-spot. Our findings suggest that RYBP might be a rescuer of the high expression of PKP1 in tumors,
where it could decrease the epithelial–mesenchymal transition in some cancer cells.},
organization = {Spanish Ministry of Economy and Competitiveness and
European ERDF Funds (MCIU/AEI/Feder, EU)},
organization = {PID2021-127296OB-I00 to A.V.-C.
and CP19/00095 to C.d.J.R},
organization = {Fondo de Investigaciones Sanitarias from Instituto de Salud Carlos
III and European Union (ERDF/ESF, “Investing in your future”)},
organization = {PI21/00394 to
O.A. and PI22/00824 to C.d.J.R.},
organization = {PAIDI program, Group BIO309},
organization = {Junta de Andalucía and Fondo de
Investigaciones Sanitarias from Instituto de Salud Carlos III, grant number PI10/00198, to M.E.F.-
V.},
organization = {Diputación General de Aragón, grant numbers E45-20R to A.V.-C. and B25-20R to O.A.},
organization = {Consellería de Innovación, Universidades, Ciencia y Sociedad Digital (Comunidad Valenciana), grant
number CIAICO 2021/0135 to J.L.N. and C.d.J.R.},
publisher = {MDPI},
keywords = {Immunofluorescence},
keywords = {Protein-protein interactions},
keywords = {Intrinsically disordered protein},
title = {Unveiling the Binding between the Armadillo-Repeat Domain of Plakophilin 1 and the Intrinsically Disordered Transcriptional Repressor RYBP},
doi = {10.3390/biom14050561},
author = {Araujo-Abad, Salome and Rizzuti, Bruno and Vidal, Miguel and Abian, Olga and Farez Vidal, María Esther and Velázquez Campoy, Adrián and De Juan Romero, Camino and Neira, José Luis},
}