000056107 001__ 56107
000056107 005__ 20210121114537.0
000056107 0247_ $$2doi$$a10.1016/j.jphotobiol.2015.07.006
000056107 0248_ $$2sideral$$a92858
000056107 037__ $$aART-2015-92858
000056107 041__ $$aeng
000056107 100__ $$aLuján, M.A.
000056107 245__ $$aIn vivo reconstitution of a homodimeric cytochrome b559 like structure: The role of the N-terminus a-subunit from Synechocystis sp. PCC 6803
000056107 260__ $$c2015
000056107 5060_ $$aAccess copy available to the general public$$fUnrestricted
000056107 5203_ $$aThe cytochrome b559 is a heme-bridged heterodimeric protein with two subunits, a and ß. Both subunits from Synechocystis sp. PCC 6803 have previously been cloned and overexpressed in Escherichia coli and in vivo reconstitution experiments have been carried out. The formation of homodimers in the bacterial membrane with endogenous heme was only observed in the case of the ß-subunit (ß/. ß) but not with the full length a-subunit. In the present work, reconstitution of a homodimer (a/. a) cytochrome b559 like structure was possible using a chimeric N-terminus a-subunit truncated before the amino acid isoleucine 17, eliminating completely a short amphipathic a-helix that lays on the surface of the membrane. Overexpression and in vivo reconstitution in the bacteria was clearly demonstrated by the brownish color of the culture pellet and the use of a commercial monoclonal antibody against the fusion protein carrier, the maltoside binding protein, and polyclonal antibodies against a synthetic peptide of the a-subunit from Thermosynechococcus elongatus. Moreover, a simple partial purification after membrane solubilization with Triton X-100 confirmed that the overexpressed protein complex corresponded with the maltoside binding protein-chimeric a-subunit cytochrome b559 like structure. The features of the new structure were determined by UV-Vis, electron paramagnetic resonance and redox potentiometric techniques. Ribbon representations of all possible structures are also shown to better understand the mechanism of the cytochrome b559 maturation in the bacterial cytoplasmic membrane.
000056107 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B18$$9info:eu-repo/grantAgreement/ES/DGA/B33$$9info:eu-repo/grantAgreement/ES/MICINN/BFU2010-16297$$9info:eu-repo/grantAgreement/ES/MINECO/AGL2011-23574$$9info:eu-repo/grantAgreement/ES/MINECO/BIO2012-35271$$9info:eu-repo/grantAgreement/ES/MINECO/MAT2011-23861
000056107 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000056107 590__ $$a3.035$$b2015
000056107 591__ $$aBIOPHYSICS$$b22 / 72 = 0.306$$c2015$$dQ2$$eT1
000056107 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b119 / 289 = 0.412$$c2015$$dQ2$$eT2
000056107 592__ $$a0.723$$b2015
000056107 593__ $$aBiophysics$$c2015$$dQ2
000056107 593__ $$aRadiology, Nuclear Medicine and Imaging$$c2015$$dQ2
000056107 593__ $$aRadiological and Ultrasound Technology$$c2015$$dQ2
000056107 593__ $$aRadiation$$c2015$$dQ2
000056107 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000056107 700__ $$0(orcid)0000-0002-5406-3280$$aMartínez, J.I.$$uUniversidad de Zaragoza
000056107 700__ $$0(orcid)0000-0003-3449-4929$$aAlonso, P.J.$$uUniversidad de Zaragoza
000056107 700__ $$aTorrado, A.
000056107 700__ $$aRoncel, M.
000056107 700__ $$aOrtega, J.M.
000056107 700__ $$0(orcid)0000-0002-2879-9200$$aSancho, J.$$uUniversidad de Zaragoza
000056107 700__ $$aPicorel, R.
000056107 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000056107 7102_ $$12003$$2395$$aUniversidad de Zaragoza$$bDpto. Física Materia Condensa.$$cÁrea Física Materia Condensada
000056107 773__ $$g152 (2015), 308–317$$pJ. photochem. photobiol., B Biol.$$tJOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY$$x1011-1344
000056107 8564_ $$s978436$$uhttps://zaguan.unizar.es/record/56107/files/texto_completo.pdf$$yPostprint
000056107 8564_ $$s80603$$uhttps://zaguan.unizar.es/record/56107/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000056107 909CO $$ooai:zaguan.unizar.es:56107$$particulos$$pdriver
000056107 951__ $$a2021-01-21-11:14:33
000056107 980__ $$aARTICLE