New applications of 2D filtered/edited NOESY for assignment and structure elucidation of RNA and RNA-protein complexes.
NMR spectra of large RNAs are difficult to assign because of extensive spectral overlap and unfavorable relaxation properties. Here we present a new approach to facilitate assignment of RNA spectra using a suite of four 2D-filtered/edited NOESY experiments in combination with base-type-specific isotopically labeled RNA. The filtering method was developed for use in 3D filtered NOESY experiments (Zwahlen et al., 1997), but the 2D versions are both more sensitive and easier to interpret for larger RNAs than their 3D counterparts. These experiments are also useful for identifying intermolecular NOEs in RNA-protein complexes. Applications to NOE assignment of larger RNAs and an RNA-protein complex are presented.
Peterson
Robert D
RD
Department of Chemistry and Biochemistry, and Molecular Biology Institute, University of California, Los Angeles, CA 90095-1569, U.S.A.
Theimer
Carla A
CA
Wu
Haihong
H
Feigon
Juli
J
D046911:Macromolecular Substances; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000295:instrumentation / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D043244:Ribonuclease III / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D019098:Telomerase / Q000737:chemistry
D046911:Macromolecular Substances; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase; C468367:RNT1 protein, S cerevisiae; D043244:Ribonuclease III