NMR studies of an FK-506 analog, [U-13C]ascomycin, bound to FK-506-binding protein. |
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Authors: | A M Petros G Gemmecker P Neri E T Olejniczak D Nettesheim R X Xu E G Gubbins H Smith S W Fesik |
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Institution: | Pharmaceutical Discovery Division, Abbott Park, Illinois 60064. |
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Abstract: | 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. |
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