Crystallization and preliminary crystallographic study of a ternary complex between the T4 phage beta-glucosyltransferase, uridine diphosphoglucose and a DNA fragment containing an abasic site.
A base-flipping phenomenon has been established for DNA methyltransferases and for DNA base-excision repair glycosylases and is likely to prove general for enzymes that need access to DNA bases to undergo chemical reaction. T4 phage beta-glucosyltransferase (BGT) is a good candidate for this novel mechanism. In order to confirm this, BGT was crystallized with an abasic site-containing DNA and uridine diphosphoglucose (UDP-glucose). The crystallization strategy is described. A complete data set was collected at 1.8 A resolution on a Cu Kalpha rotating-anode X-ray source. Molecular replacement was performed and the initial electron-density maps clearly show bound DNA.
Larivière
Laurent
L
Laboratoire d'Enzymologie et Biochimie Structurales, UPR 9063 CNRS, Bâtiment 34, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette, France.
Kurzeck
Jürgen
J
Aschke-Sonnenborn
Ursula
U
Rüger
Wolfgang
W
Moréra
Solange
S
D017122:Bacteriophage T4 / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004279:DNA, Viral / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D014532:Uridine Diphosphate Glucose / Q000737:chemistry*
D017931:DNA Primers; D004279:DNA, Viral; D005964:Glucosyltransferases; D014532:Uridine Diphosphate Glucose