title: A successful strategy for the recovering of active P21, an insoluble recombinant protein of Trypanosoma cruzi.
abstract: Structural studies of proteins normally require large quantities of pure material that can only be obtained through heterologous expression systems and recombinant technique. In these procedures, large amounts of expressed protein are often found in the insoluble fraction, making protein purification from the soluble fraction inefficient, laborious, and costly. Usually, protein refolding is avoided due to a lack of experimental assays that can validate correct folding and that can compare the conformational population to that of the soluble fraction. Herein, we propose a validation method using simple and rapid 1D (1)H nuclear magnetic resonance (NMR) spectra that can efficiently compare protein samples, including individual information of the environment of each proton in the structure.
authors.affiliation: 1] Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil [2].
authors.forename: Marlus Alves
authors.initials: MA
authors.lastname: dos Santos
authors.affiliation: 1] Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brasil [2].
authors.forename: Francesco Brugnera
authors.initials: FB
authors.lastname: Teixeira
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brasil.
authors.forename: Heline Hellen Teixeira
authors.initials: HH
authors.lastname: Moreira
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Adele Aud
authors.initials: AA
authors.lastname: Rodrigues
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Fabrício Castro
authors.initials: FC
authors.lastname: Machado
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Tatiana Mordente
authors.initials: TM
authors.lastname: Clemente
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Paula Cristina
authors.initials: PC
authors.lastname: Brigido
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Rebecca Tavares e
authors.initials: RT
authors.lastname: Silva
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Cecílio
authors.initials: C
authors.lastname: Purcino
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Rafael Gonçalves Barbosa
authors.initials: RG
authors.lastname: Gomes
authors.affiliation: 1] Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo - EPM/UNIFESP, SP, Brasil [2] Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais-ICB/UFMG, Minas Gerais, Brasil.
authors.forename: Diana
authors.initials: D
authors.lastname: Bahia
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo - EPM/UNIFESP, SP, Brasil.
authors.forename: Renato Arruda
authors.initials: RA
authors.lastname: Mortara
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brasil.
authors.forename: Claudia Elisabeth
authors.initials: CE
authors.lastname: Munte
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brasil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Horjales
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Claudio Vieira
authors.initials: CV
authors.lastname: da Silva
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D051379:Mice; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation; D058849:Protein Refolding; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D012995:Solubility; D014349:Trypanosoma cruzi / Q000235:genetics; D014709:Vero Cells
chemical_list: D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins