TY - JOUR
T1 - Studying RNA using site-directed spin-labeling and continuous-wave electron paramagnetic resonance spectroscopy.
AU - Zhang, Xiaojun
AU - Cekan, Pavol
AU - Sigurdsson, Snorri Th
AU - Qin, Peter Z.
N1 - Funding Information: This research was supported by the National Institutes of Health (R01 GM069557, PZQ), Nation Science Foundation (MCB 054652, PZQ), and the Icelandic Research Fund (60028021, STS).
PY - 2009
Y1 - 2009
N2 - In site-directed spin-labeling (SDSL), a stable nitroxide radical is attached to a specific location within a macromolecule and electron paramagnetic resonance (EPR) spectroscopy is used to interrogate the local environment surrounding the nitroxide. The SDSL strategy enables probing site-specific structural and dynamic features of RNA in solution without being limited by the size of the molecule, thus serving as a unique tool in biophysical studies of RNA. This chapter describes the use of continuous-wave (cw)-EPR to study dynamic features of RNAs as well as to monitor interactions between them. Various approaches for attaching nitroxide spin labels to nucleic acids are described, followed by detailed descriptions of cw-EPR spectral acquisition and processing procedures. Specific examples are subsequently used to illustrate analysis of EPR spectra, showing how information regarding the parent RNA can be extracted.
AB - In site-directed spin-labeling (SDSL), a stable nitroxide radical is attached to a specific location within a macromolecule and electron paramagnetic resonance (EPR) spectroscopy is used to interrogate the local environment surrounding the nitroxide. The SDSL strategy enables probing site-specific structural and dynamic features of RNA in solution without being limited by the size of the molecule, thus serving as a unique tool in biophysical studies of RNA. This chapter describes the use of continuous-wave (cw)-EPR to study dynamic features of RNAs as well as to monitor interactions between them. Various approaches for attaching nitroxide spin labels to nucleic acids are described, followed by detailed descriptions of cw-EPR spectral acquisition and processing procedures. Specific examples are subsequently used to illustrate analysis of EPR spectra, showing how information regarding the parent RNA can be extracted.
UR - https://www.scopus.com/pages/publications/77149142936
U2 - 10.1016/s0076-6879(09)69015-7
DO - 10.1016/s0076-6879(09)69015-7
M3 - Article
C2 - 20946796
SN - 0076-6879
VL - 469
SP - 303
EP - 328
JO - Methods in Enzymology
JF - Methods in Enzymology
ER -