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

Sci Rep. 2014 Mar 4:4:4259. doi: 10.1038/srep04259.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Mice
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Denaturation
  • Protein Refolding
  • 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*
  • Solubility
  • Trypanosoma cruzi* / genetics
  • Vero Cells

Substances

  • Protozoan Proteins
  • Recombinant Fusion Proteins