Summary
The solvent-exposed regions of [U-13C]ascomycin when bound to its putative target protein; FKBP, have been identified based on the different proton longitudinal relaxation rates (R1 = 1/T1) measured in the absence and presence of the paramagnetic relaxation reagent, 4-hydroxy-2,2,6,6-tetramethyl-piperidinyl-I-oxy (HyTEMPO). The proton T1s of bound ascomycin were determined using a pulse sequence (T1-HMQC) which consists of a 180° proton pulse and a variable delay (τ) followed by a heteronuclear multiple quantum correlation (HMQC) experiment. The solvent-exposed regions of ascomycin determined by these experiments are compared to NOE data in which ascomycin/FKBP contacts were identified and to the X-ray structure of the FK-506/FKBP complex.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arai T., Koyama Y., Suenaga, T. and Honda, H. (1962)J. Antibiotics,15, 231–232.
Arseniev, A.S., Sobol, A.G. and Bystrov, V.F. (1986)J. Magn. Reson.,70, 427–435.
Fesik, S. W. (1989) InComputer-Aided Drug Design. Methods and Applications (Eds, Perun, T.J. and Propst, C.L.) Marcel Dekker, New York, pp. 133–184.
Fesik, S.W., Gemmecker, G., Olejniczak, E.T. and Petros, A.M. (1991)J. Am. Chem. Soc.,113, 7080–7081.
Harding, M.W., Galat, A., Uehling, D.E. and Schreiber, S.L. (1989)Nature,341, 758–760.
Hatanaka, H., Kino, T., Miyata, S., Inamura, N., Kuroka, A., Goto, T., Tanaka, H. and Okuhara, M. (1988)J. Antibiotics,41, 1592–1601.
Maki, N., Sekiguchi, F., Nishimaki, J., Miwa, K., Hayano, T., Takahashi, N. and Suzuki, M. (1990)Proc. Natl. Acad. Sci. U.S.A.,87, 5440–5443.
Müller, L. (1979)J. Am. Chem. Soc.,101, 4481–4484.
Niccolai, N., Valensin, G., Rossi, C. and Gibbons, W.A. (1982) J. Am. Chem. Sac.,104, 1534–1537.
Petros, A.M., Müller, L. and Kopple, K.D. (1990)Biochemistry,29, 10041–10048.
Petros, A.M., Gampe, Jr., R.T., Gemmecker, G., Neri, P., Holzman, T.F., Edalji, R., Hochlowski, J., Jackson, M., McAlpine, J., Luly, J.R., Pilot-Matias, T., Pratt, S. and Fesik, S.W. (1991)J. Med. Chem.,34, 2925–2928.
Siekierka, J.J., Hung, S.H.Y., Poe, M., Lin, C.S. and Sigal, N.H. (1989)Nature,341, 755–757.
Solomon, I. and Bloembergen, J. (1956)J. Chem. Phys.,25, 261–266.
Standaert, R.F., Galat, A., Verdine, G.L. and Schreiber, S.L. (1990)Nature,46, 671–674.
Van Duyne, G.D., Standaert, R.F., Karplus, P.A., Schreiber, S.L. and Clardy, J. (1991)Science,252, 839–842.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Petros, A.M., Neri, P. & Fesik, S.W. Identification of solvent-exposed regions of an FK-506 analog, ascomycin, bound to FKBP using a paramagnetic probe. J Biomol NMR 2, 11–18 (1992). https://doi.org/10.1007/BF02192797
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02192797