Crystallization and preliminary X-ray analysis of a domain in the Runx2 transcription factor that interacts with the 1alpha,25 dihydroxy vitamin D3 receptor
Biochemistry & Molecular Pharmacology
Graduate School of Biomedical Sciences; Department of Cell Biolog
Medical Subject Headings
Absorptiometry, Photon; Animals; Core Binding Factor Alpha 1 Subunit; Crystallization; Mice; Protein Binding; Protein Structure, Tertiary; Receptors, Calcitriol
Life Sciences | Medicine and Health Sciences
The Runx2 transcription factor is a key regulator of osteoblast differentiation. In response to 1alpha,25 dihydroxy vitamin D3, Runx2 may interact with the 1alpha,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 A and a complete data set to a 3.3 A resolution was collected and analyzed. The crystals belong to the tetragonal system, with a space group P4 and unit-cell parameters of a = b = 90.8, and c = 57.2 A. The presence of a monomer of the recombinant GST-Runx2(I(209-361)) in the asymmetric unit gives a V(M) of 2.7 A(3) Da(-1) and a solvent content of 54.8%.
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Citation: J Cell Biochem. 2007 Jun 1;101(3):785-9. Link to article on publisher's site
Bruna, Carola; Arriagada, Gloria; Lian, Jane B.; Stein, Gary S.; Bunster, Marta; Martinez-Oyanedel, Jose; and Montecino, Martin A., "Crystallization and preliminary X-ray analysis of a domain in the Runx2 transcription factor that interacts with the 1alpha,25 dihydroxy vitamin D3 receptor" (2007). GSBS Student Publications. 135.