Carbon-proton scalar couplings in RNA. 3D heteronuclear and 2D isotope-edited NMR of a [sup 13]C-labeled extra-stable hairpin
- Univ. of California, Berkeley, CA (United States) Lawrence Berkeley Lab., CA (United States)
Long range carbon-proton scalar couplings were measured for an RNA hairpin of 12 nucleotides using 3D and [sup 13]C-edited 2D NMR. The large one-bond carbon-proton scalar couplings ([sup 1]J[sub CH]) and small n-bond couplings ([sup 1]J[sub CH]) produce ECOSY type cross-peaks, thus facilitating the determination of the sign and magnitude of the smaller [sup 2]J[sub CH] or [sup 3]J[sub CH]. The UUCGRNA hairpin (5[prime]-rGGACUUCGGUCC-3[prime]), whose structure has been determined by our laboratory, was uniformly [sup 13]C-labeled at 30% isotopic enrichment. The observed [sup 1]J[sub CH] couplings were then correlated to the known structure. The signs of [sup 2]J[sub C4[prime]H5[prime]], [sup 2]J[sub C4[prime]H5[double prime]], and [sup 2]J[sub C5[prime]H4[prime]], can be used for the stereospecific assignment of H5[prime](pro-S)/H5[double prime](pro-R) protons and for determining torsion angle [gamma](05[prime]-C5[prime]-C4[prime]-C3[prime]). The sign of any one of the [sup 2]J[sub C1[prime]H2[prime]], [sup 2]J[sub C2[prime]H3[prime]], [sup 2]J[sub C3[prime]H2[prime]], and [sup 2]J[sub C4[prime]H3[prime]] can be used to determine the ribose sugar conformation. Comparison of the magnitude of [sup 3]J[sub C3[prime]H5[prime]] and [sup 3]J[sub C3[prime]H5[double prime]] can also be used for the stereospecific assignment of H5[prime](pro-S)/H5[double prime](pro-R). The utility and potential limitations of using [sup 2]J[sub CH] for RNA structure determination are discussed in-depth. 38 refs., 3 figs., 5 tabs.
- DOE Contract Number:
- FG03-86ER60406; FG05-86ER75281
- OSTI ID:
- 7159832
- Journal Information:
- Journal of the American Chemical Society; (United States), Vol. 116:13; ISSN 0002-7863
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
RNA
MOLECULAR STRUCTURE
NUCLEAR MAGNETIC RESONANCE
SPECTROSCOPY
BACTERIA
CARBON 13
COMPILED DATA
COUPLING
ENZYMES
LABELLED COMPOUNDS
CARBON ISOTOPES
DATA
EVEN-ODD NUCLEI
INFORMATION
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MICROORGANISMS
NUCLEI
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC COMPOUNDS
PROTEINS
RESONANCE
STABLE ISOTOPES
550400* - Genetics
550200 - Biochemistry
400102 - Chemical & Spectral Procedures
550700 - Microbiology