000079666 001__ 79666
000079666 005__ 20191126134631.0
000079666 0247_ $$2doi$$a10.1016/j.bbagen.2018.03.009
000079666 0248_ $$2sideral$$a106432
000079666 037__ $$aART-2018-106432
000079666 041__ $$aeng
000079666 100__ $$aSantofimia-Castaño, Patricia
000079666 245__ $$aAmphipathic helical peptides hamper protein-protein interactions of the intrinsically disordered chromatin nuclear protein 1 (NUPR1)
000079666 260__ $$c2018
000079666 5060_ $$aAccess copy available to the general public$$fUnrestricted
000079666 5203_ $$aBackground: NUPR1 is a multifunctional intrinsically disordered protein (IDP) involved, among other functions, in chromatin remodelling, and development of pancreatic ductal adenocarcinoma (PDAC). It interacts with several biomolecules through hydrophobic patches around residues Ala33 and Thr68. The drug trifluoperazine (TFP), which hampers PDAC development in xenografted mice, also binds to those regions. Because of the large size of the hot-spot interface of NUPR1, small molecules could not be adequate to modulate its functions. Methods: We explored how amphipathic helical-designed peptides were capable of interacting with wild-type NUPR1 and the Thr68Gln mutant, inhibiting the interaction with NUPR1 protein partners. We used in vitro biophysical techniques (fluorescence, circular dichroism (CD), nuclear magnetic resonance (NMR) and isothermal titration calorimetry (ITC)), in silico studies (docking and molecular dynamics (MD)), and in cellulo protein ligation assays (PLAs) to study the interaction. Results: Peptide dissociation constants towards wild-type NUPR1 were ~ 3 µM, whereas no interaction was observed with the Thr68Gln mutant. Peptides interacted with wild-type NUPR1 residues around Ala33 and residues at the C terminus, as shown by NMR. The computational results clarified the main determinants of the interactions, providing a mechanism for the ligand-capture that explains why peptide binding was not observed for Thr68Gln mutant. Finally, the in cellulo assays indicated that two out of four peptides inhibited the interaction of NUPR1 with the C-terminal region of the Polycomb RING protein 1 (C-RING1B). Conclusions: Designed peptides can be used as lead compounds to inhibit NUPR1 interactions. General significance: Peptides may be exploited as drugs to target IDPs.
000079666 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B01$$9info:eu-repo/grantAgreement/ES/DGA/B89$$9info:eu-repo/grantAgreement/ES/FIS/PI15-00663$$9info:eu-repo/grantAgreement/ES/ISCIII/CPII13-0017$$9info:eu-repo/grantAgreement/ES/MINECO/BFU2013-47064-P$$9info:eu-repo/grantAgreement/ES/MINECO/BFU2016-78232-P$$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-64445-R
000079666 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000079666 590__ $$a3.681$$b2018
000079666 591__ $$aBIOPHYSICS$$b19 / 72 = 0.264$$c2018$$dQ2$$eT1
000079666 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b103 / 294 = 0.35$$c2018$$dQ2$$eT2
000079666 592__ $$a1.455$$b2018
000079666 593__ $$aBiochemistry$$c2018$$dQ1
000079666 593__ $$aMolecular Biology$$c2018$$dQ1
000079666 593__ $$aBiophysics$$c2018$$dQ1
000079666 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000079666 700__ $$aRizzuti, Bruno
000079666 700__ $$0(orcid)0000-0001-5664-1729$$aAbián, Olga$$uUniversidad de Zaragoza
000079666 700__ $$0(orcid)0000-0001-5702-4538$$aVelázquez-Campoy, Adrián$$uUniversidad de Zaragoza
000079666 700__ $$aIovanna, Juan L.
000079666 700__ $$aNeira, J. L.
000079666 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000079666 773__ $$g1862, 6 (2018), 1283-1295$$pBiochim. biophys. acta (G)$$tBIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS$$x0304-4165
000079666 8564_ $$s918625$$uhttps://zaguan.unizar.es/record/79666/files/texto_completo.pdf$$yPostprint
000079666 8564_ $$s70079$$uhttps://zaguan.unizar.es/record/79666/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000079666 909CO $$ooai:zaguan.unizar.es:79666$$particulos$$pdriver
000079666 951__ $$a2019-11-26-13:41:26
000079666 980__ $$aARTICLE