Expression and purification of untagged Glnk proteins from actinobacteria

Authors
Gerhardt, Edileusa C. M.; Moure, Vivian R.; Souza, Andrey W.; Pedrosa, Fabio O.; Souza, Emanuel M.; Diacovich, Lautaro; Gramajo, Hugo Cesar; Huergo, Luciano F.
Publication Year
2017
Language
English
Format
article
Status
Published version
Description
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 welldocumented 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 efficientpurification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtrationanalysis 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.
Fil: Gerhardt, Edileusa C. M.. Universidade Federal do Paraná; Brasil
Fil: Moure, Vivian R.. Universidade Federal do Paraná; Brasil
Fil: Souza, Andrey W.. Universidade Federal do Paraná; Brasil
Fil: Pedrosa, Fabio O.. Universidade Federal do Paraná; Brasil
Fil: Souza, Emanuel M.. Universidade Federal do Paraná; Brasil
Fil: Diacovich, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Huergo, Luciano F.. Universidade Federal do Paraná; Brasil
Subject
PII PROTEINS
ACTINOBACTERIA
NITROGEN
METABOLISM
Otras Ciencias Biológicas
Ciencias Biológicas
CIENCIAS NATURALES Y EXACTAS
Access level
Open access
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/53368