Crystals of trp repressor suitable for high-resolution neutron Laue diffraction studies.
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 A were recorded from a D(2)O-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.
Daniels
Brenda V
BV
Biology Department, Brookhaven National Laboratory, Upton, NY 11973, USA.
Myles
Dean A A
DA
Forsyth
V Trevor
VT
Lawson
Catherine L
CL
D001426:Bacterial Proteins; D003460:Crystallization / Q000379:methods; D004268:DNA-Binding Proteins / Q000737:chemistry; D003903:Deuterium; D004058:Diffusion; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007532:Isoleucine / Q000235:genetics; D033363:Neutron Diffraction / Q000379:methods; D012097:Repressor Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D012997:Solvents / Q000737:chemistry; D014633:Valine / Q000235:genetics; D029968:Escherichia coli Proteins
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D003903:Deuterium; D007532:Isoleucine; D012097:Repressor Proteins; D012997:Solvents; D014633:Valine; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins