title: Use of a fusion protein to obtain crystals suitable for X-ray analysis: crystallization of a GST-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF.
abstract: Crystals of glutathione-S-transferase (GST)-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF, were obtained under crystallization conditions similar to those for GST. Preliminary X-ray crystallographic analysis revealed that crystals of the GST-fused protein belong to space group P6(1)22 or P6(5)22 with unit cell dimensions a = b = 140.4 A, c = 93.5 A and gamma = 120 degrees, having one molecule in the crystallographic asymmetric unit. The crystals diffract to 2.5 A resolution. The cell dimensions are related to those of GST crystals thus far reported. Crystallization of the DNA-binding domain that was cleaved from the fused protein by thrombin was also carried out using several methods under numerous conditions, but efforts to produce well-ordered large crystals were unsuccessful. A possible application of GST-fusion proteins for small target proteins or domains to obtain crystals suitable for X-ray structure determination is proposed.
authors.affiliation: Department of Molecular Biology, Nara Institute of Science and Technology (NAIST), Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kuge
authors.forename: Y
authors.initials: Y
authors.lastname: Fujii
authors.forename: T
authors.initials: T
authors.lastname: Shimizu
authors.forename: F
authors.initials: F
authors.lastname: Hirose
authors.forename: A
authors.initials: A
authors.lastname: Matsukage
authors.forename: T
authors.initials: T
authors.lastname: Hakoshima
mesh_terms: D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005982:Glutathione Transferase / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
chemical_list: D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase