Crystallization and preliminary X-ray analysis of the RAD protein from Antirrhinum majus.
Crystals of the RADIALIS protein from Antirrhinum majus were grown by vapour diffusion after limited proteolysis. Mass spectrometry indicated that an 8 kDa fragment had been crystallized corresponding to the predicted MYB DNA-binding domain. X-ray data collected at room temperature were consistent with tetragonal symmetry, whereas data collected at 100 K using crystals cryoprotected by supplementing the mother liquor with ethylene glycol conformed to orthorhombic symmetry. It was subsequently shown that crystals soaked in cryoprotectants that were ;osmolality-matched' to the mother liquor retained tetragonal symmetry. Using these crystals, X-ray data were collected in-house to a maximum resolution of 2 A.
Stevenson
Clare E M
CE
Department of Biological Chemistry, John Innes Centre, Norwich NR4 7UH, England.
Burton
Nicolas
N
Costa
Manuela
M
Nath
Utpal
U
Dixon
Ray A
RA
Coen
Enrico S
ES
Lawson
David M
DM
D032161:Antirrhinum / Q000378:metabolism*; D015925:Cryopreservation; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D019855:Ethylene Glycol / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013696:Temperature; D014157:Transcription Factors / Q000737:chemistry*; D014961:X-Ray Diffraction
D014157:Transcription Factors; D004247:DNA; D019855:Ethylene Glycol