PLoS ONE 2015-01-01

Purification and characterization of a DNA-binding recombinant PREP1:PBX1 complex.

Lisa Mathiasen, Chiara Bruckmann, Sebastiano Pasqualato, Francesco Blasi

Index: PLoS ONE 10(4) , e0125789, (2015)

Full Text: HTML

Abstract

Human PREP1 and PBX1 are homeodomain transcriptional factors, whose biochemical and structural characterization has not yet been fully described. Expression of full-length recombinant PREP1 (47.6 kDa) and PBX1 (46.6 kDa) in E. coli is difficult because of poor yield, high instability and insufficient purity, in particular for structural studies. We cloned the cDNA of both proteins into a dicistronic vector containing an N-terminal glutathione S-transferase (GST) tag and co-expressed and co-purified a stable PBX1:PREP1 complex. For structural studies, we produced two C-terminally truncated complexes that retain their ability to bind DNA and are more stable than the full-length proteins through various purification steps. Here we report the production of large amounts of soluble and pure recombinant human PBX1:PREP1 complex in an active form capable of binding DNA.


Related Compounds

Related Articles:

Salicylic acid signaling controls the maturation and localization of the arabidopsis defense protein ACCELERATED CELL DEATH6.

2014-08-01

[Mol. Plant 7(8) , 1365-83, (2014)]

Transcriptional regulation of Munc13-4 expression in cytotoxic lymphocytes is disrupted by an intronic mutation associated with a primary immunodeficiency.

2014-06-02

[J. Exp. Med. 211(6) , 1079-91, (2014)]

Irisin stimulates muscle growth-related genes and regulates adipocyte differentiation and metabolism in humans.

2012-07-01

[Int. J. Obes. 38(12) , 1538-44, (2014)]

Epigenetic reprogramming of the type III interferon response potentiates antiviral activity and suppresses tumor growth.

2014-01-01

[PLoS Biol. 12(1) , e1001758, (2014)]

Mechanism of human PTEN localization revealed by heterologous expression in Dictyostelium.

2014-12-11

[Oncogene 33(50) , 5688-96, (2014)]

More Articles...