151021 20251017144622.0 doi 10.1016/j.abb.2022.109125 sideral 128385 ART-2022-128385 eng Neira, José L. Biochemical and biophysical characterization of PADI4 supports its involvement in cancer 2022 Access copy available to the general public Unrestricted PADI4 (protein-arginine deiminase, also known as protein L-arginine iminohydrolase) is one of the human isoforms of a family of Ca2+-dependent proteins catalyzing the conversion of arginine to citrulline. Although the consequences of this process, known as citrullination, are not fully understood, all PADIs have been suggested to play essential roles in development and cell differentiation. They have been found in a wide range of cells and tissues and, among them, PADI4 is present in macrophages, monocytes, granulocytes and cancer cells. In this work, we focused on the biophysical features of PADI4 and, more importantly, how its expression was altered in cancer cells. Firstly, we described the different expression patterns of PADI4 in various cancer cell lines and its colocalization with the tumor-related protein p53. Secondly, we carried out a biophysical characterization of PADI4, by using a combination of biophysical techniques and in silico molecular dynamics simulations. Our biochemical results suggest the presence of several forms of PADI4 with different subcellular localizations, depending on the cancer cell line. Furthermore, PADI4 could have a major role in tumorigenesis by regulating p53 expression in certain cancer cell lines. On the other hand, the native structure of PADI4 was strongly pH-dependent both in the absence or presence of Ca2+, and showed two pH-titrations at basic and acidic pH values. Thus, there was a narrow pH range (from 6.5 to 8.0) where the protein was dimeric and had a native structure, supporting its role in histones citrullination. Thermal denaturations were always two-state, but guanidinium-induced ones showed that PADI4 unfolded through at least one intermediate. Our simulation results suggest that the thermal melting of PADI4 structure was rather homogenous throughout its sequence. The overall results are discussed in terms of the functional role of PADI4 in the development of cancer. info:eu-repo/grantAgreement/ES/DGA/B25-20R info:eu-repo/grantAgreement/ES/DGA/E45-20R info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI18-00349-Investing in your future info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/BFU2016-78232-P info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00 info:eu-repo/grantAgreement/ES/MINECO/CP19-00095 info:eu-repo/semantics/openAccess by-nc-nd https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es 3.9 2022 BIOPHYSICS 17 / 70 = 0.243 2022 Q1 T1 BIOCHEMISTRY & MOLECULAR BIOLOGY 126 / 285 = 0.442 2022 Q2 T2 0.867 2022 Biophysics 2022 Q1 Molecular Biology 2022 Q2 Biochemistry 2022 Q2 7.5 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Araujo-Abad, Salomé Cámara-Artigas, Ana Rizzuti, Bruno Abian, Olga Universidad de Zaragoza (orcid)0000-0001-5664-1729 Giudici, Ana Marcela Velazquez-Campoy, Adrián Universidad de Zaragoza (orcid)0000-0001-5702-4538 Juan Romero, Camino de 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 717 (2022), 109125 [16 pp.] Arch. biochem. biophys. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 0003-9861 2825199 http://zaguan.unizar.es/record/151021/files/texto_completo.pdf Versión publicada 2451183 http://zaguan.unizar.es/record/151021/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:151021 articulos driver 2025-10-17-14:22:11 ARTICLE