Isotope-detected /sup 1/H NMR studies of proteins: a general strategy for editing interproton nuclear Overhauser effects by heteronuclear decoupling, with application to phage lambda repressor
A strategy for editing interproton nuclear Overhauser effects (NOEs) in proteins is proposed and illustrated. Selective incorporation of /sup 13/C- (or /sup 15/N)-labeled amino acids into a protein permits NOEs involving the labeled residues to be identified by heteronuclear difference decoupling. Such heteronuclear editing simplifies the NOE difference spectrum and avoids ambiguities due to spin diffusion. Isotope-detected /sup 1/H NMR thus opens to study proteins too large for conventional one- and two-dimensional NMR methods (20-75 kDa). The authors have applied this strategy to the N-terminal domain of phage lambda repressor, a protein of dimer molecular mass 23 kDa. A tertiary NOE from an internal aromatic ring (Phe-51) to a ..beta..-/sup 13/C-labeled alanine residue (Ala-62) is demonstrated.
- Research Organization:
- Harvard Univ., Cambridge, MA (United States)
- OSTI ID:
- 5559180
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:5
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GENE REPRESSORS
MOLECULAR STRUCTURE
NUCLEAR MAGNETIC RESONANCE
ALANINES
AMINO ACIDS
BACTERIOPHAGES
CARBON 13
ESCHERICHIA COLI
NITROGEN 15
OVERHAUSER EFFECT
PROTEINS
PROTONS
BACTERIA
BARYONS
CARBON ISOTOPES
CARBOXYLIC ACIDS
ELEMENTARY PARTICLES
EVEN-ODD NUCLEI
FERMIONS
HADRONS
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MICROORGANISMS
NITROGEN ISOTOPES
NUCLEI
NUCLEONS
NUCLEOPROTEINS
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PARASITES
RESONANCE
STABLE ISOTOPES
VIRUSES
550601* - Medicine- Unsealed Radionuclides in Diagnostics