000060970 001__ 60970
000060970 005__ 20190709135501.0
000060970 0247_ $$2doi$$a10.1038/srep43880
000060970 0248_ $$2sideral$$a98517
000060970 037__ $$aART-2017-98517
000060970 041__ $$aeng
000060970 100__ $$aHuijbers, M. M. E.
000060970 245__ $$aProline dehydrogenase from Thermus thermophilus does not discriminate between FAD and FMN as cofactor
000060970 260__ $$c2017
000060970 5060_ $$aAccess copy available to the general public$$fUnrestricted
000060970 5203_ $$aFlavoenzymes are versatile biocatalysts containing either FAD or FMN as cofactor. FAD often binds to a Rossmann fold, while FMN prefers a TIM-barrel or flavodoxin-like fold. Proline dehydrogenase is denoted as an exception: it possesses a TIM barrel-like fold while binding FAD. Using a riboflavin auxotrophic Escherichia coli strain and maltose-binding protein as solubility tag, we produced the apoprotein of Thermus thermophilus ProDH (MBP-TtProDH). Remarkably, reconstitution with FAD or FMN revealed that MBP-TtProDH has no preference for either of the two prosthetic groups. Kinetic parameters of both holo forms are similar, as are the dissociation constants for FAD and FMN release. Furthermore, we show that the holo form of MBP-TtProDH, as produced in E. coli TOP10 cells, contains about three times more FMN than FAD. In line with this flavin content, the crystal structure of TtProDH variant ¿ABC, which lacks helices aA, aB and aC, shows no electron density for an AMP moiety of the cofactor. To the best of our knowledge, this is the first example of a flavoenzyme that does not discriminate between FAD and FMN as cofactor. Therefore, classification of TtProDH as an FAD-binding enzyme should be reconsidered.
000060970 536__ $$9info:eu-repo/grantAgreement/ES/MICINN/BIO2013-42978-P
000060970 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000060970 590__ $$a4.122$$b2017
000060970 591__ $$aMULTIDISCIPLINARY SCIENCES$$b12 / 64 = 0.188$$c2017$$dQ1$$eT1
000060970 592__ $$a1.533$$b2017
000060970 593__ $$aMultidisciplinary$$c2017$$dQ1
000060970 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000060970 700__ $$0(orcid)0000-0001-9047-0046$$aMartínez-Júlvez, M.$$uUniversidad de Zaragoza
000060970 700__ $$aWestphal, A. H.
000060970 700__ $$aDelgado-Arciniega, E.
000060970 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, M.$$uUniversidad de Zaragoza
000060970 700__ $$aVan Berkel, W. J. H.
000060970 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000060970 773__ $$g7 (2017), 43880 [13 pp.]$$pSci. rep.$$tScientific reports$$x2045-2322
000060970 8564_ $$s1365790$$uhttps://zaguan.unizar.es/record/60970/files/texto_completo.pdf$$yVersión publicada
000060970 8564_ $$s110249$$uhttps://zaguan.unizar.es/record/60970/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000060970 909CO $$ooai:zaguan.unizar.es:60970$$particulos$$pdriver
000060970 951__ $$a2019-07-09-11:46:34
000060970 980__ $$aARTICLE