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

Laboratoire d'Enzymologie et Biochimie Structurales, UPR 9063 CNRS, Bâtiment 34, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette, France.

Published: September 2002

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.

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http://dx.doi.org/10.1107/S0907444902012969DOI Listing

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