A successful strategy for the recovering of active P21, an insoluble recombinant protein of Trypanosoma cruzi

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 purificatio...

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Published in:	Scientific reports Vol. 4; no. 1; p. 4259
Main Authors:	Santos, Marlus Alves dos, Teixeira, Francesco Brugnera, Moreira, Heline Hellen Teixeira, Rodrigues, Adele Aud, Machado, Fabrício Castro, Clemente, Tatiana Mordente, Brigido, Paula Cristina, Silva, Rebecca Tavares e., Purcino, Cecílio, Gomes, Rafael Gonçalves Barbosa, Bahia, Diana, Mortara, Renato Arruda, Munte, Claudia Elisabeth, Horjales, Eduardo, da Silva, Claudio Vieira
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 04-03-2014 Nature Publishing Group
Subjects:	101/6 14/63 140/131 38 631/326/417/1716 631/57/2272/2273 82/80 82/83 Animals Cercopithecus aethiops Escherichia coli - genetics Escherichia coli - metabolism GTP-binding protein Humanities and Social Sciences Mice multidisciplinary NMR Nuclear magnetic resonance Nuclear Magnetic Resonance, Biomolecular Protein Denaturation Protein folding Protein purification Protein Refolding Proteins Protozoa Protozoan Proteins - chemistry Protozoan Proteins - genetics Protozoan Proteins - isolation & purification Protozoan Proteins - metabolism Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - isolation & purification Recombinant Fusion Proteins - metabolism Science Solubility Trypanosoma cruzi - genetics Vero Cells
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2045-2322 2045-2322
DOI:	10.1038/srep04259