000136154 001__ 136154
000136154 005__ 20240719195438.0
000136154 0247_ $$2doi$$a10.1016/j.bpc.2024.107288
000136154 0248_ $$2sideral$$a139165
000136154 037__ $$aART-2024-139165
000136154 041__ $$aeng
000136154 100__ $$aNeira, José L.
000136154 245__ $$aIsolated auto-citrullinated regions of PADI4 associate to the intact protein without altering their disordered conformation
000136154 260__ $$c2024
000136154 5060_ $$aAccess copy available to the general public$$fUnrestricted
000136154 5203_ $$aPADI4 is one of the human isoforms of a group of enzymes intervening in the conversion of arginine to citrulline. It is involved in the development of several types of tumors, as well as other immunological illnesses, such as psoriasis, multiple sclerosis, or rheumatoid arthritis. PADI4 auto-citrullinates in several regions of its sequence, namely in correspondence of residues Arg205, Arg212, Arg218, and Arg383. We wanted to study whether the citrullinated moiety affects the conformation of nearby regions and its binding to intact PADI4. We designed two series of synthetic peptides comprising either the wild-type or the relative citrullinated versions of such regions – i.e., a first series of peptides comprising the first three arginines, and a second series comprising Arg383. We studied their conformational properties in isolation by using fluorescence, far-ultraviolet (UV) circular dichroism (CD), and 2Dsingle bond1H NMR. Furthermore, we characterized the binding of the wild-type and citrullinated peptides in the two series to the intact PADI4, by using isothermal titration calorimetry (ITC), fluorescence, and biolayer interferometry (BLI), as well as by molecular docking simulations. We observed that citrullination did not alter the local conformational propensities of the isolated peptides. Nevertheless, for all the peptides in the two series, citrullination slowed down the kinetic koff rates of the binding reaction to PADI4, probably due to differences in electrostatic effects compared to the presence of arginine. The affinities of PADI4 for unmodified peptides were slightly larger than those of the corresponding citrullinated ones in the two series, but they were all within the same range, indicating that there were no relevant variations in the thermodynamics of binding due to sequence effects. These results highlight details of the self-citrullination of PADI4 and, more generally, of possible auto-catalytic mechanisms taking place in vivo for other citrullinating enzymes or, alternatively, in proteins undergoing citrullination passively.
000136154 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B25-23R$$9info:eu-repo/grantAgreement/ES/DGA/E45-23R$$9info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI21-00394$$9info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-I00
000136154 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000136154 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000136154 700__ $$aRizzuti, Bruno
000136154 700__ $$0(orcid)0000-0001-5664-1729$$aAbian, Olga$$uUniversidad de Zaragoza
000136154 700__ $$0(orcid)0000-0001-5702-4538$$aVelazquez-Campoy, Adrian$$uUniversidad de Zaragoza
000136154 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000136154 773__ $$g312 (2024), 107288 [12 pp.]$$pBiophys. chemist.$$tBiophysical Chemistry$$x0301-4622
000136154 8564_ $$s5086164$$uhttps://zaguan.unizar.es/record/136154/files/texto_completo.pdf$$yVersión publicada
000136154 8564_ $$s2512983$$uhttps://zaguan.unizar.es/record/136154/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000136154 909CO $$ooai:zaguan.unizar.es:136154$$particulos$$pdriver
000136154 951__ $$a2024-07-19-18:28:17
000136154 980__ $$aARTICLE