Crystallization and preliminary X-ray analysis of a domain in the Runx2 transcription factor that interacts with the 1alpha,25 dihydroxy vitamin D3 receptor
Lian, Jane B.
Stein, Gary S.
Montecino, Martin A.
UMass Chan AffiliationsDepartment of Cell Biolog
Graduate School of Biomedical Sciences
Document TypeJournal Article
KeywordsAbsorptiometry, Photon; Animals; Core Binding Factor Alpha 1 Subunit; Crystallization; Mice; Protein Binding; Protein Structure, Tertiary; Receptors, Calcitriol
Medicine and Health Sciences
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AbstractThe 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%.
SourceJ Cell Biochem. 2007 Jun 1;101(3):785-9. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/32797
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