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

Author(s):Show less -

dos Santos, Marlus Alves ^[1] ; Teixeira, Francesco Brugnera ^[2] ; Teixeira Moreira, Heline Hellen ^[2] ; Rodrigues, Adele Aud ^[1] ; Machado, Fabricio Castro ^[1] ; Clemente, Tatiana Mordente ^[1] ; Brigido, Paula Cristina ^[1] ; Silva, Rebecca Tavares E. ^[1] ; Purcino, Cecilio ^[1] ; Barbosa Gomes, Rafael Goncalves ^[1] ; Bahia, Diana ^{[3, 4]} ; Mortara, Renato Arruda ^[4] ; Munte, Claudia Elisabeth ^[2] ; Horjales, Eduardo ^[2] ; da Silva, Claudio Vieira ^[1]

Affiliation

^[1] Univ Fed Uberlandia, Inst Ciencias Biomed, BR-38400 Uberlandia, MG - Brazil

^[2] Univ Sao Paulo, Inst Fis Sao Carlos, Sao Carlos, SP - Brazil

^[3] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Biol Geral, Belo Horizonte, MG - Brazil

^[4] Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Vila Mariana, SP - Brazil

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 4, MAR 4 2014.
• Web of Science Citations: 4

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 H-1 nuclear magnetic resonance (NMR) spectra that can efficiently compare protein samples, including individual information of the environment of each proton in the structure. (AU)

• FAPESP's process: 12/21153-7 - Structural studies on Trypanosoma cruzi P21 protein as a strategy for Chagas Disease treatment
• Grantee: Francesco Brugnera Teixeira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 10/51867-6 - SMOLBNET 2.0: Structural studies on proteins related to trypanosomatids infection and to complexes of these proteins with molecules which are involved in the infection process
• Grantee: Eduardo Horjales Reboredo
• Support Opportunities: Regular Research Grants