/sup 32/P and 2-dimensional NMR methodology for assignment of /sup 31/P and /sup 2/H signals of oligonucleotides
Because of spectral overlap, even with 2-D methods, /sup 1/H signal assignments in oligonucleotides much longer than tetramers is exceedingly difficult. However, by chemically introducing site-specific 0-17, 0-18, and S labeling in the phosphoryl groups of oligonucleotides, it is now possible to unambiguously assign the /sup 32/P peaks. Thus, it is possible to assign all three phosphate /sup 31/P signals of the oligonucleotide tetramer d(ApGpCpT) by site-specific introduction of the three different oxygen isotopes into the three different phosphate diesters. Using two-dimensional /sup 31/P//sup 1/H correlated spectral methods the authors can also unambiguously identify the /sup 1/H NMR signals coupled to the assigned /sup 31/P signals. In the latter, only those protons which are scalar coupled to the /sup 31/P nucleus are observed in the 2-D heteronuclear spectrum. Finally by the /sup 1/H//sup 1/H COSY and /sup 1/H NOESY they can identify the other protons of the oligonucleotides. This new methodology is not dependent upon any assumed B-DNA structure as is required in other recent 2-D oligonucleotide assignment techniques. Assignment of signals in the actinomycin D intercalating d(ApGpCpT) tetramer complex, d(CGCAGAATTCGCG), and lac operator pseudo-fragment, d(TGTGAGCGCTCACA), are now possible.
- Research Organization:
- Purdue Univ., West Lafayette, IN
- OSTI ID:
- 5097257
- Report Number(s):
- CONF-8606151-; TRN: 86-034823
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
NUCLEOTIDES
MOLECULAR STRUCTURE
NMR SPECTRA
DEUTERIUM
NUCLEAR MAGNETIC RESONANCE
PHOSPHORUS 32
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
HYDROGEN ISOTOPES
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RESONANCE
SPECTRA
STABLE ISOTOPES
550201* - Biochemistry- Tracer Techniques
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