000061946 001__ 61946
000061946 005__ 20190709135505.0
000061946 0247_ $$2doi$$a10.1038/s41598-017-07875-5
000061946 0248_ $$2sideral$$a100706
000061946 037__ $$aART-2017-100706
000061946 041__ $$aeng
000061946 100__ $$0(orcid)0000-0001-7202-4587$$aSebastián, M.$$uUniversidad de Zaragoza
000061946 245__ $$aKinetics and thermodynamics of the protein-ligand interactions in the riboflavin kinase activity of the FAD synthetase from Corynebacterium ammoniagenes
000061946 260__ $$c2017
000061946 5060_ $$aAccess copy available to the general public$$fUnrestricted
000061946 5203_ $$aEnzymes known as bifunctional and bimodular prokaryotic type-I FAD synthetase (FADS) exhibit ATP:riboflavin kinase (RFK) and FMN:ATP adenylyltransferase (FMNAT) activities in their C-Terminal and N-Terminal modules, respectively, and produce flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). These act as cofactors of a plethora of flavoproteins in all organisms. Therefore, regulation of their production maintains the cellular flavoproteome homeostasis. Here, we focus on regulation of the FMN synthesis in Corynebacterium ammoniagenes (Ca) by the inhibition of its RFK activity by substrates and products of the reaction. We use a truncated CaFADS variant consisting in the isolated C-Terminal RFK module, whose RFK activity is similar to that of the full-length enzyme. Inhibition of the RFK activity by the RF substrate is independent of the FMNAT module, and FMN production, in addition to being inhibited by an excess of RF, is also inhibited by both of the reaction products. Pre-steady-state kinetic and thermodynamic studies reveal key aspects to the substrates induced fit to produce the catalytically competent complex. Among them, the role of Mg2+ in the concerted allocation of substrates for catalysis and the ensemble of non-competent complexes that contribute to the regulated inhibition of the RFK activity are particularly relevant.
000061946 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B18$$9info:eu-repo/grantAgreement/ES/MINECO/BIO2013-42978-P$$9info:eu-repo/grantAgreement/ES/MINECO/BIO2016-75183-P
000061946 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000061946 590__ $$a4.122$$b2017
000061946 591__ $$aMULTIDISCIPLINARY SCIENCES$$b12 / 64 = 0.188$$c2017$$dQ1$$eT1
000061946 592__ $$a1.533$$b2017
000061946 593__ $$aMultidisciplinary$$c2017$$dQ1
000061946 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000061946 700__ $$aSerrano, A.$$uUniversidad de Zaragoza
000061946 700__ $$0(orcid)0000-0001-5702-4538$$aVelázquez-Campoy, A.$$uUniversidad de Zaragoza
000061946 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, M.$$uUniversidad de Zaragoza
000061946 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000061946 773__ $$g7 (2017), [14 pp.]$$pSci. rep.$$tScientific reports$$x2045-2322
000061946 8564_ $$s3106924$$uhttps://zaguan.unizar.es/record/61946/files/texto_completo.pdf$$yVersión publicada
000061946 8564_ $$s111362$$uhttps://zaguan.unizar.es/record/61946/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000061946 909CO $$ooai:zaguan.unizar.es:61946$$particulos$$pdriver
000061946 951__ $$a2019-07-09-11:48:12
000061946 980__ $$aARTICLE