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Crystallization and preliminary X‐ray analysis of a domain in the Runx2 transcription factor that interacts with the 1α,25 dihydroxy vitamin D3 receptor
Author(s) -
Bruna Carola,
Arriagada Gloria,
Lian Jane B.,
Stein Gary S.,
Bunster Marta,
MartinezOyanedel Jose,
Montecino Martin
Publication year - 2007
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.21231
Subject(s) - transcription factor , crystallization , chemistry , domain (mathematical analysis) , calcitriol receptor , runx2 , microbiology and biotechnology , receptor , biology , biochemistry , mathematics , gene , mathematical analysis , organic chemistry
The Runx2 transcription factor is a key regulator of osteoblast differentiation. In response to 1α,25 dihydroxy vitamin D3, Runx2 may interact with the 1α,25 dihydroxy vitamin D3 receptor (VDR) in the promoter of target genes, producing a synergic activation of their transcription. Previous studies have suggested that the motifs responsible for the VDR–Runx2 interaction are contained within the 230–361 domain of Runx2. In this work, we confirmed by GST‐pull down that Runx2 I(209–361) is sufficient to interact with the VDR. To obtain structural information, GST–Runx2 I(209–361) protein was overexpressed in Escherichia coli , purified and crystallized using the hanging‐drop vapor‐diffusion method and polyethyleneglycol as a precipitant. The crystals were found to diffract to a maximum resolution of 2.7 Å and a complete data set to a 3.3 Å resolution was collected and analyzed. The crystals belong to the tetragonal system, with a space group P 4 and unit‐cell parameters of a = b = 90.8, and c = 57.2 Å. The presence of a monomer of the recombinant GST–Runx2 I(209–361) in the asymmetric unit gives a V M of 2.7 Å 3 Da −1 and a solvent content of 54.8%. J. Cell. Biochem. 101: 785–789, 2007. © 2007 Wiley‐Liss, Inc.