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

Acta Crystallogr D Biol Crystallogr. 2002 Sep;58(Pt 9):1484-6. doi: 10.1107/S0907444902012969. Epub 2002 Aug 23.

Abstract

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.

MeSH terms

  • Bacteriophage T4 / enzymology*
  • Bacteriophage T4 / genetics
  • Base Sequence
  • Crystallization
  • Crystallography, X-Ray
  • DNA Primers
  • DNA, Viral / chemistry*
  • Electrophoretic Mobility Shift Assay
  • Glucosyltransferases / chemistry*
  • Glucosyltransferases / isolation & purification
  • Protein Conformation
  • Uridine Diphosphate Glucose / chemistry*

Substances

  • DNA Primers
  • DNA, Viral
  • Glucosyltransferases
  • Uridine Diphosphate Glucose