Selective deuteration of RNA for NMR signal assignment.
For site-selective deuterium labeling of RNA, [5-2H]uridine phosphoramidite was prepared. The uridine at position 10 of a 25-mer RNA, GGACAGACUUCGGUCGGAGUACUCG, was labeled in two different manners for "positive" (U = [5-1H]U, U = [5-2H] U) and "negative"(U = [5-2H]U, U = [5-1H]U) observations. By comparison of NOESY spectra of the two labeled samples with that of the unlabeled RNA, we could unambiguously assign the H5-H6 signals of U10, and measure their NOE connectivities.
Kim
I
I
Department of Biophysics and Biochemistry, School of Science, University of Tokyo, Japan.
Watanabe
S
S
Muto
Y
Y
Hosono
K
K
Takai
K
K
Takaku
H
H
Kawai
G
G
Watanabe
K
K
Yokoyama
S
S
D001483:Base Sequence; D001665:Binding Sites; D003903:Deuterium / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D011131:Polyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry*
D011131:Polyribonucleotides; D012313:RNA; D006859:Hydrogen; D003903:Deuterium