Comparison of alignment tensors generated for native tRNA(Val) using magnetic fields and liquid crystalline media.
Residual dipolar couplings (RDCs) complement standard NOE distance and J-coupling torsion angle data to improve the local and global structure of biomolecules in solution. One powerful application of RDCs is for domain orientation studies, which are especially valuable for structural studies of nucleic acids, where the local structure of a double helix is readily modeled and the orientations of the helical domains can then be determined from RDC data. However, RDCs obtained from only one alignment media generally result in degenerate solutions for the orientation of multiple domains. In protein systems, different alignment media are typically used to eliminate this orientational degeneracy, where the combination of RDCs from two (or more) independent alignment tensors can be used to overcome this degeneracy. It is demonstrated here for native E. coli tRNA(Val) that many of the commonly used liquid crystalline alignment media result in very similar alignment tensors, which do not eliminate the 4-fold degeneracy for orienting the two helical domains in tRNA. The intrinsic magnetic susceptibility anisotropy (MSA) of the nucleobases in tRNA(Val) was also used to obtain RDCs for magnetic alignment at 800 and 900 MHz. While these RDCs yield a different alignment tensor, the specific orientation of this tensor combined with the high rhombicity for the tensors in the liquid crystalline media only eliminates two of the four degenerate orientations for tRNA(Val). Simulations are used to show that, in optimal cases, the combination of RDCs obtained from liquid crystalline medium and MSA-induced alignment can be used to obtain a unique orientation for the two helical domains in tRNA(Val).
Latham
Michael P
MP
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, CO 80309-0215, USA.
Hanson
Paul
P
Brown
Darin J
DJ
Pardi
Arthur
A
D016880:Anisotropy; D001483:Base Sequence; D004926:Escherichia coli / Q000737:chemistry*; D050866:Liquid Crystals / Q000737:chemistry*; D008280:Magnetics; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D025341:Principal Component Analysis; D012366:RNA, Transfer, Val / Q000737:chemistry* / Q000235:genetics
D009587:Nitrogen Isotopes; D012366:RNA, Transfer, Val