Exprssion and purification of untagged GlnK proteins from Actinobacteria

EDILEUSA C.M. GERHARDT, VIVIAN R. MOURE, ANDREY W. SOUZA, FABIO O. PEDROSA, EMANUEL M. SOUZA, LAUTARO DIACOVICH, HUGO GRAMAJO, LUCIANO F. HUERGO

EXCLI JOURNAL

UNIV MAINZ-MED DEPT

Año: 2017

1611-2156

The PII protein family constitutes one of the most conserved and well distributed family of signal transductionproteins 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 andfunction 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 willfacilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria