Ultra-High Field NMR Studies of Stable Isotope Applications
EMSL Project ID
2438a
Abstract
The primary goal of this project is to develop and implement new approaches for using isotopic labels in NMR-based DNA structure determination that will allow highly accurate NMR-based DNA structure determination for damaged DNAs, longer DNAs in solution (20-30 bp) and DNAs in complex with proteins. This is a DOE funded collaboration between the stable isotopes group at LANL and the NMR scientists at PNNL. Historically, NMR-based DNA structure determination has been an under-restrained problem. Isotopic-labeling can considerably increase the number and type of NMR experiments that can be collected on DNA samples resulting in more accurate structures. However, uniform or specific labeling of synthetic DNA oligomers has not yet become widespread due to intrinsic difficulty and cost. In our project, the primary approach is to generate site specifically-labeled samples using phosphoramidite chemistry, an approach that is expensive and challenging, but that allows maximum flexibility in labeling, in comparison to enzymatic or pcr-based labeling approaches.
Project Details
Project type
Capability Research
Start Date
2003-04-01
End Date
2006-04-02
Status
Closed
Released Data Link
Team
Principal Investigator
Related Publications
McAteer K, A Aceves Gaona, R Michalczyk, GW Buchko, NG Isern, LA Silks, JH Miller, and MA Kennedy. 2004. "Compensating Bends in a 16 base-pair DNA Oligomer Containing a T3A3 Segment: A NMR Study of Global DNA Curvature." Biopolymers 75(6):497-511.