Expression and purification of untagged GlnK proteins from actinobacteria

Authors

  • Edileusa C.M. Gerhardt Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil
  • Vivian R. Moure Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil
  • Andrey W. Souza Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil
  • Fabio O. Pedrosa Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil
  • Emanuel M. Souza Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil
  • Lautaro Diacovich Instituto de Biologia Molecular y Celular de Rosario (IBR-CONICET), Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Rosario, Argentina
  • Hugo Gramajo Instituto de Biologia Molecular y Celular de Rosario (IBR-CONICET), Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Rosario, Argentina
  • Luciano F. Huergo Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba, Brazil; Setor Litoral, UFPR, Matinhos, Brazil

DOI:

https://doi.org/10.17179/excli2017-394

Keywords:

PII, GlnK, CD analysis, actinobacteria

Abstract

The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been well documented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficient purification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtration analysis supported that the purified proteins are correctly folded. The purification protocol described here will facilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria.

Published

2017-06-27

How to Cite

Gerhardt, E. C., Moure, V. R., Souza, A. W., Pedrosa, F. O., Souza, E. M., Diacovich, L., … Huergo, L. F. (2017). Expression and purification of untagged GlnK proteins from actinobacteria . EXCLI Journal, 16, 949–958. https://doi.org/10.17179/excli2017-394

Issue

Section

Original articles