Crystals of trp repressor suitable for high-resolution neutron Laue diffraction studies

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 59; no. 1; pp. 136 - 138
• Main Authors: Daniels, Brenda V., Myles, Dean A. A., Forsyth, V. Trevor, Lawson, Catherine L.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.01.2003
• Subjects: Bacterial Proteins; Crystallization - methods; Deuterium; Diffusion; DNA recognition; DNA-Binding Proteins - chemistry; Escherichia coli - chemistry; Escherichia coli - genetics; Escherichia coli - metabolism; Isoleucine - genetics; Laue; neutron diffraction; Neutron Diffraction - methods; Repressor Proteins - biosynthesis; Repressor Proteins - chemistry; Repressor Proteins - genetics; Solvents - chemistry; trp repressor; Valine - genetics
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S0907444902018516

Crystallization and preliminary neutron‐diffraction measurements of wild‐type variant Val58→Ile of the Escherichia coli trp repressor are reported. A vapor‐diffusion chamber suitable for initial protein‐solution volumes in the range 0.2–0.5 ml was used to grow cube‐shaped crystals with edge dimensions in the range 0.8–1.4 mm. Neutron Laue measurements to a nominal resolution of 2.1 Å were recorded from a D2O‐exchanged crystal using the LADI instrument at ILL. These results demonstrate that it will be possible for the first time to obtain a full‐atom neutron structural model of a DNA‐binding protein plus its associated solvent. Direct observation of hydrogen bonding between protein and solvent should enhance understanding of the role of solvent in protein–DNA recognition.