Investigation of the role of Mg2+ in DNA repair proteins APE1, Pol ?, and FEN1
EMSL Project ID
10600c
Abstract
We propose to utilize unique capabilities within the EMSL (low temperature solid-state NMR and the mpp2 super computer) to investigate Mg2+ binding to two DNA repair proteins Pol ? and APE1. In the case of Pol ? we are interested in the order of binding of Mg2+ with respect to the binding of damaged DNA. The binding of Mg2+ to Pol ? in the absence of DNA could explain the high error rate of this polymerase. In the case of APE1, the Mg2+ NMR experiments will resolve a key mechanistic question relative to APE1, i.e. does it operate via a single- or two-metal mechanism. To perform and analyze the results of these experiments we need access to 12 weeks (2016 hours) of wide bore 500 time and a similar amount of time on the 400 wide bore spectrometer. For analysis of the results we need 75,000 cpu-hours of mpp2 time.
Project Details
Project type
Capability Research
Start Date
2006-04-06
End Date
2007-05-21
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Lipton AS, RW Heck, SV Primak, DR McNeill, DM Wilson Iii, and PD Ellis. 2008. "Characterization of Mg2+ Binding to the DNA Repair Protein Apurinic/Apyrimidic Endonuclease 1 via Solid-State 25Mg NMR Spectroscopy." Journal of the American Chemical Society 130(29):9332-9341. doi:10.1021/ja0776881