| Autor: |
Stockman BJ; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115., Nirmala NR, Wagner G, Delcamp TJ, DeYarman MT, Freisheim JH |
| Jazyk: |
angličtina |
| Zdroj: |
Biochemistry [Biochemistry] 1992 Jan 14; Vol. 31 (1), pp. 218-29. |
| DOI: |
10.1021/bi00116a031 |
| Abstrakt: |
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 15N was required to obtain the resonance assignments via heteronuclear 3D NMR spectroscopy since homonuclear 2D spectra did not provide sufficient 1H resonance dispersion. Medium- and long-range NOE's have been used to characterize the secondary structure of the binary ligand-enzyme complex in solution. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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