1WVL

Crystal Structure of Multimeric DNA-binding Protein Sac7d-GCN4 with DNA decamer

Summary for 1WVL

• Entry DOI: 10.2210/pdb1wvl/pdb
• Related: 1AZP 1WD0
• Descriptor: 5'-D(*CP*CP*TP*AP*TP*AP*TP*AP*GP*G)-3', DNA-binding proteins 7a/7b/7d, GCN4 (3 entities in total)
• Functional Keywords: leucine zipper, protein engineering, synchrotron radiation, thermophile, structural protein-dna complex, structural protein/dna
• Biological source: Sulfolobus acidocaldarius
• Total number of polymer chains: 4
• Total formula weight: 24525.65

Authors

Wu, S.W.,Wang, A.H. (deposition date: 2004-12-16, release date: 2005-08-02, Last modification date: 2023-10-25)

Primary citation

Wu, S.W.,Ko, T.P.,Chou, C.C.,Wang, A.H.
Design and characterization of a multimeric DNA binding protein using Sac7d and GCN4 as templates
Proteins, 60:617-628, 2005

PubMed Abstract: The protein Sac7d belongs to a class of small chromosomal proteins from the hyperthermophilic archaeon Sulfolobus acidocaldarius. Sac7d is extremely stable to heat, acid, and chemical agents. This protein is a monomer and it binds DNA without any particular sequence preference, while inducing a sharp kink in the DNA. By appending a leucine-zipper-like helical peptide derived from the yeast transcriptional activator GCN4 to the C-terminal end of Sac7d, the modified monomers (denoted S7dLZ) are expected to interact with each other via hydrophobic force to form a parallel dimer. The recombinant S7dLZ was expressed in Escherichia coli and purified by heating and ion-exchange chromatography. The formation of dimer was detected by gel-filtration chromatography and chemical cross-link. The results of surface plasmon resonance and circular dichroism experiments showed that the DNA-binding capacity was retained. Furthermore, X-ray diffraction analysis of single crystals of S7dLZ in complex with DNA decamer CCTATATAGG showed that the leucine-zipper segments of S7dLZ were associated into an antiparallel four-helix bundle. There are two DNA fragments bound to each S7dLZ tetramer in the crystal. This model works as a successful template that endows protein a new function without losing original properties.

PubMed: 16028219
DOI: 10.1002/prot.20524

Experimental method

X-RAY DIFFRACTION (2.6 Å)