NMR studies of an FK-506 analog, [U-13C]ascomycin, bound to FK-506-binding protein.

Petros AM, Gemmecker G, Neri P, Olejniczak ET, Nettesheim D, Xu RX, Gubbins EG, Smith H, Fesik SW
J Med Chem. 1992 35 (13): 2467-73

PMID: 1377749 · DOI:10.1021/jm00091a015

Multidimensional, heteronuclear NMR methods were used to determine the complete 1H and 13C resonance assignments for [U-13C]ascomycin bound to recombinant FKBP, including stereospecific assignment of all 22 methylene protons. The conformation of ascomycin was then determined from an analysis of NOEs observed in a 13C-edited 3D HMQC-NOESY spectrum of the [U-13C]ascomycin/FKBP. This structure is found to be quite different from the solution structure of the two forms of uncomplexed FK-506. However, it is very similar to the X-ray crystal structure of FK-506 bound to FKBP, rms deviation = 0.56 A. The methods used for resonance assignment and structure calculation are presented in detail. Furthermore, FKBP/ascomycin NOEs are reported which help define the structure of the ascomycin binding pocket. This structural information obtained in solution was compared to the recently described X-ray crystal structure of the FKBP/FK-506 complex.

MeSH Terms (13)

Carbon Isotopes Carrier Proteins Cell Line Escherichia coli Gene Expression Genes, Bacterial Humans Magnetic Resonance Spectroscopy Recombinant Proteins T-Lymphocytes Tacrolimus Tacrolimus Binding Proteins X-Ray Diffraction

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