## Chemistry Department Faculty Citations

#### Title

Crystallization of Escherichia coli catabolite gene activator protein with its DNA binding site: The use of modular DNA

#### ACS Citation

Schultz, S. C.; Shields, G. C.; Steitz, T. A. Crystallization of Escherichia coli Catabolite Gene Activator Protein with Its DNA Binding Site: The Use of Modular DNA. J. Mol. Biol. 1990, 213 (1), 159-166.

#### Abstract

To obtain crystals of the Escherichia coli catabolite gene activator protein (CAP) complexed with its DNA-binding site, we have searched for crystallization conditions with 26 different DNA segments â‰¥28 base-pairs in length that explore a variety of nucleotide sequences, lengths, and extended 5â€² or 3â€² termini. In addition to utilizing uninterrupted asymmetric lac site sequences, we devised a novel approach of synthesizing half-sites that allowed us to efficiently generate symmetric DNA segments with a wide variety of extended termini and lengths in the large size range (â‰¥28 bp) required by this protein. We report three crystal forms that are suitable for X-ray analysis, one of which (crystal form III) gives measurable diffraction amplitudes to 3 {\AA} resolution. Additives such as calcium, n-octyl-$\beta$-d-glucopyranoside and spermine produce modest improvements in the quality of diffraction from crystal form III. Adequate stabilization of crystal form III is unexpectedly complex, requiring a greater than tenfold reduction in the salt concentration followed by addition of 2-methyl-2,4-pentanediol and then an increase in the concentration of polyethylene glycol.

#### Source Name

Journal of Molecular Biology

1990

213

1

159-166

Citation

Article

COinS