London Research Institute
-Clare Hall Laboratories-
-Cancer Research UK-
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Molecular Enzymology
London Research Institute -Clare Hall Laboratories- -Cancer Research UK- Charity Registration No. 1089464 -Terms and Conditions- |
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| Crystallisation and Structural Determination |
The DNA substrate we decided to use was a model of a stalled fork in which synthesis on the lagging strand had proceeded slightly further than on the leading strand (figure 2). Biochemical studies2 have shown that this is the preferred substrate for RecG.
Figure 2. Schematic diagram of the DNA substrate RecG was co-crystallised with. The arms of the fork are identified as template (the unreplicated duplex), leading (the parental leading strand) and lagging (the newly synthesised daughter strand). The length of each arm in base-pairs are shown in pink. Note the one base-pair gap in the lagging strand at the junction. The structure of the enzyme from the hyperthermophile Thermotoga maritima was determined to 3.2Å, and comprised the RecG protein, the bound three-way fork, and a bound ADP molecule and magnesium ion. The coordinates have been deposited in the RCSB Protein Data Bank under identification code 1GM5. |