Sequence-specific sup 1 H and sup 15 N resonance assignments for human dihydrofolate reductase in solution

Journal Article · Tue Jan 14 00:00:00 EST 1992 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00116a031· OSTI ID:5688133

Stockman, B J; Nirmala, N R; Wagner, G ^[1]; Delcamp, T J; DeYarman, M T; Freisheim, J H ^[2]

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1. Harvard Medical School, Boston, MA (United States)
2. Medical Coll. of Ohio, Toledo (United States)

Dihydrofolate reductase is an intracellular target enzyme for folate antagonists, including the anticancer drug methotrexate. In order to design novel drugs with altered binding properties, a detailed description of protein-drug interactions in solution is desirable to understand the specificity of drug binding. As a first step in this process, heteronuclear three-dimensional NMR spectroscopy has been used to make sequential resonance assignments for more than 90% of the residues in human dihydrofolate reductase complexed with methotrexate. Uniform enrichment of the 21.5-kDa protein with {sup 15}N was required to obtain the resonance assignments via heteronuclear 3D NMR spectroscopy since homonuclear 2D spectra did not provide sufficient {sup 1}H resonance dispersion. Medium- and long-range NOE's have been used to characterize the secondary structure of the binary ligand-enzyme complex in solution.

• OSTI ID: 5688133
• Journal Information: Biochemistry; (United States), Vol. 31:1; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
OXIDOREDUCTASES
NUCLEAR MAGNETIC RESONANCE
CHEMICAL SHIFT
FOLIC ACID
MAN
MOLECULAR STRUCTURE
NITROGEN 15
OVERHAUSER EFFECT
PROTONS
AMINO ACIDS
ANIMALS
AROMATICS
AZAARENES
BARYONS
CARBOXYLIC ACIDS
DRUGS
ELEMENTARY PARTICLES
ENZYMES
FERMIONS
HADRONS
HEMATINICS
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MAMMALS
NITROGEN ISOTOPES
NUCLEI
NUCLEONS
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PRIMATES
PROTEINS
PTERIDINES
RESONANCE
STABLE ISOTOPES
VERTEBRATES
VITAMIN B GROUP
VITAMINS
550201* - Biochemistry- Tracer Techniques