Abstract
Our previous investigation of the solution structure of Fe(II)-bleomycin pointed toward the carbamoyl group in the mannose moiety or a water molecule as possible alternative axial ligands to the metal center in this metallo-bleomycin. The possibility of a solvent molecule occupying the apical position trans to the primary amine has not been ruled out yet. In order to explore this possibility even further, the coordination chemistry of azide-bound Fe(II)-bleomycin was investigated with the use of NMR applied to paramagnetic molecules. Fe(II)- and apo-bleomycin were also re-visited. Comparison of the NMR results for both Fe(II)-bound molecules obtained in the present study strongly suggests that the carbamoyl oxygen is ligated to Fe(II), and it is released from coordination upon azide binding. This event is suggested based on the diminished paramagnetic character exhibited by the carbohydrate moiety in Fe(II)-azide-bleomycin when compared with its parent metal complex. A possible structural role for the glucopyranose fragment, which changes throughout the process that starts with metallo-bleomycin formation and ends with DNA binding, is discussed. The study of the coordination of azide by Fe(II)-bleomycin through NMR has not been reported previously. Unlike magnetic CD data, NMR offers a residue-by-residue account of the possible structural changes that take place in Fe(II)-bleomycin after azide binding.
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References
Umezawa, H., Maeda, K., Takeuchi, T. & Okami, Y. New antibiotics bleomycin A and B. J. Antibiot. 19, 200–209 (1966).
Bennett, J. M. & Reich, S. D. Bleomycin. Ann. Inter. Med. 90, 945–948 (1979).
Carlson, R. W., Sikic, B. I., Turbow, M. M. & Ballon, S. C. Combination cisplatin, vinblastine, and bleomycin chemotherapy (PVB) for malignant germ-cell tumors of the ovary. J. Clin. Oncol. 1, 645–651 (1983).
Bleomycin Chemotherapy (eds Sikic, B. I., Rozencweig, M., & Carter, S. K.) Academic Press, Orlando, FL, 1985.
Einhorn, L. H. & Donohue, J. Cis-diamminedichloroplatinum, vinblastine, and bleomycin combination chemotherapy in disseminated testicular cancer. Ann. Inter. Med. 87, 293–298 (1977).
Sausville, E. A., Peisach, J. & Horwitz, S. B. Role for ferrous ion and oxygen in degradation of DNA by bleomycin. Biochem. Biophys. Res. Commun. 73, 814–822 (1976).
Hecht, S. M. RNA degradation by bleomycin, a naturally-occurring bioconjugate. Bioconjugate Chem. 5, 513–526 (1994).
Kane, S. A. & Hecht, S. M. Polynucleotide recognition and degradation by bleomycin. Prog. Nucleic Acid Res. Mol. Biol. 49, 313–352 (1994).
Hecht, S. M. Bleomycin: new perspectives on the mechanism of action. J. Nat. Prod. 63, 158–168 (2000).
Chen, J. Y. & Stubbe, J. Bleomycins: towards better therapeutics. Nat. Rev Cancer 5, 102–112 (2005).
Boger, D. L. & Cai, H. Bleomycin: synthetic and mechanistic studies. Angew. Chem. Int. Ed. 38, 448–476 (1999).
Carter, B. J., Murty, V. S., Reddy, K. S., Wang, S. -N. & Hecht, S. M. A role for the metal-binding domain in determining the DNA-sequence selectivity of Fe-bleomycin. J. Biol. Chem. 265, 4193–4196 (1990).
Hecht, S. M. The chemistry of activated bleomycin. Acc. Chem. Res. 19, 383–391 (1986).
Stubbe, J. & Kozarich, J. W. Mechanisms of bleomycin-induced DNA-degradation. Chem. Rev. 87, 1107–1136 (1987).
Povirk, L. F., Hogan, M. & Dattagupta, N. Binding of bleomycin to DNA - intercalation of the bithiazole rings. Biochemistry 18, 96–101 (1979).
Zuber, G., Quada, J. C. & Hecht, S. M. Sequence selective cleavage of a DNA octanucleotide by chlorinated bithiazoles and bleomycins. J. Am. Chem. Soc. 120, 9368–9369 (1998).
Loeb, K. E., Zaleski, J. M., Hess, C. D., Hecht, S. M. & Solomon, E. I. Spectroscopic investigation of the metal ligation and reactivity of the ferrous active sites of bleomycin and bleomycin derivatives. J. Am. Chem. Soc. 120, 1249–1259 (1998).
Lehmann, T. E. Molecular modeling of the three-dimensional structure of Fe(II)-bleomycin: are the Co(II) and Fe(II) adducts isostructural? J. Biol. Inorg. Chem. 7, 305–312 (2002).
Lehmann, T. E., Serrano, M. L. & Que, L. Jr. Coordination chemistry of Co(II)-bleomycin: Its investigation through NMR and molecular dynamics. Biochemistry 39, 3886–3898 (2000).
Akkerman, M. A. J., Neijman, E.W.J.F., Wijmenga, S. S., Hilbers, C. W. & Bermel, W. Studies of the solution structure of the bleomycin-A2 iron(II) carbon-monoxide complex by means of 2-dimensional NMR-spectroscopy and distance geometry calculations. J. Am. Chem. Soc. 112, 7462–7474 (1990).
Akkerman, M. A. J., Haasnoot, C. A. G., Pandit, U. K. & Hilbers, C. W. Complete assignment of the C-13 NMR-spectra of bleomycin-A2 and its zinc complex by means of two-dimensional NMR-spectroscopy. Magn. Reson. Chem. 26, 793–802 (1988).
Akkerman, M. A. J., Haasnoot, C. A. G. & Hilbers, C. W. Studies of the solution structure of the bleomycin-A2 zinc complex by means of two-dimensional NMR-spectroscopy and distance geometry calculations. Eur. J. Biochem. 173, 211–225 (1988).
Oppenheimer, N. J., Rodriguez, L. O. & Hecht, S. M. Structural studies of active complex of bleomycin - assignment of ligands to the ferrous ion in a ferrous-bleomycin carbon monoxide complex. PNAS 76, 5616–5620 (1979).
Lehmann, T. E., Ming, L.- J., Rosen, M. E. & Que, L. Jr. NMR studies of the paramagnetic complex Fe(II)-bleomycin. Biochemistry 36, 2807–2816 (1997).
Claussen, C. A. & Long, E. C. Nucleic acid recognition by metal complexes of bleomycin. Chem. Rev. 99, 2797–2816 (1999).
Iitaka, Y. et al. Chemistry of bleomycin 20. X-ray structure determination of P-3A Cu(II)-complex, a biosynthetic intermediate of bleomycin. J. Antibiot. 31, 1070–1072 (1978).
Masanori, S., Takanori, K., Minoru, H., Masafumi, M. & Yasuyuki, M. The 1,6 Å crystal structure of the copper(II)-bound bleomycin complexed with the bleomycin-binding protein from bleomycin-producing Streptomyces verticillus. J. Biol. Chem. 277, 2311–2320 (2002).
Goodwin, K. D., Lewis, M. A., Long, E. C. & Georgiadis, M. M. Crystal structure of DNA-bound Co(III)-bleomycin B-2: Insights on intercalation and minor groove binding. PNAS. 105, 5052–5056 (2008).
Chen, D. M., Hawkins, B. L. & Glickson, J. D. Proton nuclear magnetic-resonance study of bleomycin in aqueous-solution - assignment of resonances. Biochemistry 16, 2731–2738 (1977).
Bertini, I., Luchinat, C. & Parigi, G. Solution NMR of Paramagnetic Molecules. Applications to Metallobiomolecules and Models, in Current Methods in Inorganic Chemistry, Vol. 2 (Elsevier, New York, 2001).
Ming, L. –J. in Nuclear magnetic resonance of paramagnetic metal centers in proteins and synthetic complexes in physical methods in bioinorganic chemistry. Spectroscopy and Magnetism (ed Que, L., Jr.) 375–464 University Science Books, Sausalito, California, 2000.
Long, E. C., Georgiadis, M. M., Goodwin, K. D. & Lewis, M. A. in Bioinorganic Chemistry: Cellular Systems & Synthetic Models. ACS Symp. Series Vol. 1012 (eds Long, E. C. & Baldwin M. J.) 63–80 (ACS, Washington, D. C., 2009).
Nehaus, D. & Williamson, M. in The Nuclear Overhauser Effect in Structural and Conformational Analysis (VCH Publishers, New York, 1989).
Nagayama, K., Wüthrich, K. & Ernst, R. R. 2-Dimensional spin-echo correlated spectroscopy (SECSY) for H-1-NMR studies of biological macromolecules. Biochem. Biophys. Res. Commun. 90, 305–311 (1979).
Rance, M. et al. Improved spectral resolution in COSY H1-NMR spectra of proteins via double quantum filtering. Biochem. Biophys. Res. Commun. 117, 479–485 (1983).
Braunschweiler, L. & Ernst, R. R. Coherence transfer by isotropic mixing - application to proton correlation spectroscopy. J. Magn. Reson. 53, 521–528 (1983).
Bax, A. & Davis, D. G. MLEV-17-based two-dimensional homonuclear magnetization transfer spectroscopy. J. Magn. Reson. 65, 355–360 (1985).
Delaglio, F. et al. NMRPipe - a multidimensional spectral processing system based on Unix pipes. J. Biomol. NMR 6, 277–293 (1995).
Johnson, B. A. & Blevins, R. A. NMR view - a computer-program for the visualization and analysis of NMR data. J. Biomol. NMR 4, 603–614 (1994).
Acknowledgements
This work was supported by the University of Wyoming start-up funds. We thank Dr Douglas Wheeler (University of Wyoming, Department of Chemistry) for his assistant with some of the NMR spectra collected during this investigation. Our gratitude also goes to Dr Vladimir Alvarado (University of Wyoming, Department of Petroleum and Chemical Engineering), for reviewing this manuscript. Our gratitude also goes to Dr Bruce A Johnson (Merck Research Laboratories, Rahway, NJ, USA) and Dr Frank Delaglio (Software Science Consultant, North Potomac, MD, USA) for kindly providing the software NMRPipe and NMRView, respectively.
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Lehmann, T., Li, Y. Possible structural role of the disaccharide unit in Fe-bleomycin before and after oxygen activation. J Antibiot 65, 25–33 (2012). https://doi.org/10.1038/ja.2011.103
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DOI: https://doi.org/10.1038/ja.2011.103
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