000125955 001__ 125955
000125955 005__ 20241125101134.0
000125955 0247_ $$2doi$$a10.3389/fmolb.2023.1167348
000125955 0248_ $$2sideral$$a133557
000125955 037__ $$aART-2023-133557
000125955 041__ $$aeng
000125955 100__ $$aRivero, Maribel$$uUniversidad de Zaragoza
000125955 245__ $$aRiboflavin kinase and pyridoxine 5'-phosphate oxidase complex formation envisages transient interactions for FMN cofactor delivery
000125955 260__ $$c2023
000125955 5060_ $$aAccess copy available to the general public$$fUnrestricted
000125955 5203_ $$aEnzymes catalysing sequential reactions have developed different mechanisms to control the transport and flux of reactants and intermediates along metabolic pathways, which usually involve direct transfer of metabolites from an enzyme to the next one in a cascade reaction. Despite the fact that metabolite or substrate channelling has been widely studied for reactant molecules, such information is seldom available for cofactors in general, and for flavins in particular. Flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) act as cofactors in flavoproteins and flavoenzymes involved in a wide range of physiologically relevant processes in all type of organisms. Homo sapiens riboflavin kinase (RFK) catalyses the biosynthesis of the flavin mononucleotide cofactor, and might directly interplay with its flavin client apo-proteins prior to the cofactor transfer. Non-etheless, none of such complexes has been characterized at molecular or atomic level so far. Here, we particularly evaluate the interaction of riboflavin kinase with one of its potential FMN clients, pyridoxine-5′-phosphate oxidase (PNPOx). The interaction capacity of both proteins is assessed by using isothermal titration calorimetry, a methodology that allows to determine dissociation constants for interaction in the micromolar range (in agreement with the expected transient nature of the interaction). Moreover, we show that; i) both proteins become thermally stabilized upon mutual interaction, ii) the tightly bound FMN product can be transferred from RFK to the apo-form of PNPOx producing an efficient enzyme, and iii) the presence of the apo-form of PNPOx slightly enhances RFK catalytic efficiency. Finally, we also show a computational study to predict likely RFK-PNPOx binding modes that can envisage coupling between the FMN binding cavities of both proteins for the potential transfer of FMN.
000125955 536__ $$9info:eu-repo/grantAgreement/ES/DGA/E35-20R$$9info:eu-repo/grantAgreement/ES/DGA/LMP13_21$$9info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00
000125955 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000125955 590__ $$a3.9$$b2023
000125955 592__ $$a1.232$$b2023
000125955 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b106 / 313 = 0.339$$c2023$$dQ2$$eT2
000125955 593__ $$aBiochemistry$$c2023$$dQ1
000125955 593__ $$aBiochemistry, Genetics and Molecular Biology (miscellaneous)$$c2023$$dQ1
000125955 593__ $$aMolecular Biology$$c2023$$dQ2
000125955 594__ $$a7.2$$b2023
000125955 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000125955 700__ $$aBoneta, Sergio$$uUniversidad de Zaragoza
000125955 700__ $$aNovo, Nerea$$uUniversidad de Zaragoza
000125955 700__ $$0(orcid)0000-0001-5702-4538$$aVelázquez-Campoy, Adrián$$uUniversidad de Zaragoza
000125955 700__ $$0(orcid)0000-0001-5823-7965$$aPolo, Victor$$uUniversidad de Zaragoza
000125955 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, Milagros$$uUniversidad de Zaragoza
000125955 7102_ $$12012$$2755$$aUniversidad de Zaragoza$$bDpto. Química Física$$cÁrea Química Física
000125955 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000125955 773__ $$g10 (2023), 1167348 [19 pp.]$$pFront. mol. biosci.$$tFrontiers in Molecular Biosciences$$x2296-889X
000125955 8564_ $$s4586257$$uhttps://zaguan.unizar.es/record/125955/files/texto_completo.pdf$$yVersión publicada
000125955 8564_ $$s2119943$$uhttps://zaguan.unizar.es/record/125955/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000125955 909CO $$ooai:zaguan.unizar.es:125955$$particulos$$pdriver
000125955 951__ $$a2024-11-22-11:59:54
000125955 980__ $$aARTICLE