Human enzyme PADI4 binds to the nuclear carrier Importin a3

Neira, J. L. ; Rizzuti, B. ; Abián, O. (Universidad de Zaragoza) ; Araujo-Abad, S. ; Velázquez-Campoy, A. (Universidad de Zaragoza) ; Juan Romero, C. de

Resumen: PADI4 is a peptidyl-arginine deiminase (PADI) involved in the conversion of arginine to citrulline. PADI4 is present in macrophages, monocytes, granulocytes, and several cancer cells. It is the only PADI family member observed within both the nucleus and the cytoplasm. PADI4 has a predicted nuclear localization sequence (NLS) comprising residues Pro56 to Ser83, to allow for nuclear translocation. Recent predictors also suggest that the region Arg495 to Ile526 is a possible NLS. To understand how PADI4 is involved in cancer, we studied the ability of intact PADI4 to bind importin a3 (Impa3), a nuclear transport factor that plays tumor-promoting roles in several cancers, and its truncated species (¿Impa3) without the importin-binding domain (IBB), by using fluorescence, circular dichroism (CD), and isothermal titration calorimetry (ITC). Furthermore, the binding of two peptides, encompassing the first and the second NLS regions, was also studied using the same methods and molecular docking simulations. PADI4 interacted with both importin species, with affinity constants of ~1–5 µM. The isolated peptides also interacted with both importins. The molecular simulations predict that the anchoring of both peptides takes place in the major binding site of Impa3 for the NLS of cargo proteins. These findings suggest that both NLS regions were essentially responsible for the binding of PADI4 to the two importin species. Our data are discussed within the framework of a cell mechanism of nuclear transport that is crucial in cancer.

Idioma: Inglés

DOI: 10.3390/cells11142166

Año: 2022

Publicado en: Cells 11, 14 (2022), 2166 [28 pp.]
ISSN: 2073-4409

Factor impacto JCR: 6.0 (2022)

Categ. JCR: CELL BIOLOGY rank: 60 / 191 = 0.314 (2022) - Q2 - T1

Factor impacto CITESCORE: 9.0 - Medicine (Q1)

Factor impacto SCIMAGO: 1.537 - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B25-20R
Financiación: info:eu-repo/grantAgreement/ES/DGA/E45-20R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/PI18-0394
Financiación: info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BFU2016-78232-P
Financiación: info:eu-repo/grantAgreement/ES/MINECO/CP19-00095

Tipo y forma: Article (Published version)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)