Abstract
The proton NMR spectrum of the ternary complex between the octamer duplex d(TTGGCCAA)2, two molecules of the drug chromomycin-A3, and a divalent cobalt ion has been assigned. Assignment procedures used standard two-dimensional techniques and relied upon the expected NOE contacts observed in the equivalent diamagnetic complex containing zinc. The magnetic susceptibility tensor for the cobalt was determined and used to calculate shifts for all nuclei, aiding in the assignment process and verification. Relaxation, susceptibility, temperature and field dependence studies of the paramagnetic spectrum enabled determination of electronic properties of the octahedral cobalt complex. The electronic relaxation rate τs was determined to be 2.5 ± 1.5 ps; the effective isotropic g value was found to be 2.6 ± 0.2, indicating strong spin-orbit coupling. The magnetic susceptibility tensor was determined to be χxx = 8.9 * 10-3 cm3/mol, χyy = 9.5 * 10-3 cm3/mol, χzz = 12.8 * 10-3 cm3/mol. A tentative rotational correlation time of 8 ns was obtained for the complex. Both macroscopic and microscopic susceptibility measurements revealed deviations from Curie behavior over the temperature range accessible in the study. Non-selective relaxation rates were found to be inaccurate for defining distances from the metal center. However, pseudocontact shifts could be calculated with high accuracy using the dipolar shift equation. Isotropic hyperfine shifts were factored into contact and dipolar terms, revealing that the dipolar shift predominates and that contact shifts are relatively small.
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Banci, L., Dugad, L.B., La Mar, G.N., Keating, K. A., Luchinat, C. and Pierattelli, R. (1992) Biophys. J., 63, 530–43.
Banci, L., Bertini, I. Pierattelli, R., Tien, M. and Vila, A (1995) J. Am. Chem. Soc., 117, 8659–8667.
Banci, L., Bertini, I., Gray, H., Luchinat, C., Reddig, T., Rosato, A. and Turano, P. (1997a) Biochemistry, 36, 9867–77.
Banci, L., Bertini, I, Savellini, G.G., Romagnoli, A., Turano, P., Cremonini, M.A., Luchinat, C. and Gray, H.B. (1997b). Proteins Struct. Funct. Genet., 29, 68–76.
Banville, D.L., Keniry, M.A., Kam, M. and Shafer, R.H. (1990) Biochemistry, 29, 6521–34.
Bertini, I. and Luchinat, C. (1984). Adv. Inorg. Biochem., 6, 71–111.
Bertini, I. and Luchinat, C. (1986) In NMR of Paramagnetic Molecules in Biological Systems, Benjamin/Cummings.
Bertini, I., Luchinat, C. and Tarchi, D. (1993) Chem. Phys. Lett., 203, 445–449.
Bertini, I., Jonsson, B., Luchinat, C., Pierattelli, R. and Vila, A. (1994) J. Magn. Reson. Ser. B, 104, 230–239.
Bertini, I., Luchinat, C. and Rosato, A. (1996) Prog. Biophys. Mol. Biol., 66, 43–80.
Bertini, I., Donaire, A., Felli, I. and Rosato, A. (1996) Magn. Reson. Chem., 34, 948–50
Bertini, I., Donaire, A., Luchinat, C. and Rosato, A. (1997) Proteins, 29, 348–58.
Brown, S., Weber, P. and Müller, L. (1988) J. Magn. Reson., 77, 166.
Cheng, H. and Markley, J. (1995) Annu. Rev. Biophys. Biomol. Struct., 24, 209–237.
Cotton, F. and Wilkinson, G. (1972) In Advanced Inorganic Chemistry, John Wiley and Sons, Inc.
Day, M. (1994) STRIKER. University of California, San Francisco.
Desvaux, H. and Gochin, M., in preparation.
Emerson, S. D. and La Mar, G.N. (1990) Biochemistry, 29, 1556–66.
Ferrin, T., Huang, C., Jarvis, L. and Langridge, R. (1988). J. Mol. Graphics 6, 13–27.
Gao, X. and Patel, D.J. (1989) Biochemistry, 28, 751–762.
Gao, X. L. and Patel, D.J. (1990) Biochemistry 29, 10940–10956.
Gao, X. L., Mirau, P. and Patel, D.J. (1992) J. Mol. Biol., 223, 259–79.
Gochin, M. and Roder, H (1995) Protein Sci., 4, 296–305.
Gochin, M. (1997) J. Am. Chem. Soc., 119, 3377–3378.
Goddard, T. and Kneller, D. (1997) SPARKY. University of California, San Francisco
Goff, H. and La Mar, G.N. (1977) J. Am. Chem. Soc., 99, 6599–6606.
Goff, H. (1981) J. Am. Chem. Soc., 103, 3714–3722.
Golding, R. and Stubbs, L. (1979) J. Magn. Reson., 33, 627–647.
Griswold, B., Humoller, F. and McIntyre, A. (1951) Anal. Chem., 23, 192–194.
Harper, L. V., Amann, B. T., Kilfoil Vinson, V. and Berg, J. M. (1993) J. Am. Chem. Soc., 115, 2577–2580.
Kurland, R. and McGarvey, B. (1970) J. Magn. Reson. 2, 286–301.
La Mar, G.N., Horrocks Jr., W. and Holm, R. (1973) In NMR of Paramagnetic Molecules: Principles and Applications, Academic Press.
Levitt, M., Frenkiel, T. and Freeman, R. (1982) J. Magn. Reson., 47, 328.
Marion, D. (1994) Biochimie, 76, 631–640.
Phillips, W. and Poe, M. (1972) Methods Enzymol., 24, 304–311.
Piotto, M., Saudek, V. and Sklenar, V.J. (1992) J. Biomol. NMR, 2, 661.
Press, W., Teukolsky, S., Vetterling, W. and Flannary, B. (1992) Numerical Recipes in Fortran; The Art of Scientific Computing Cambridge University Press.
Qin, J., Delaglio, F., La Mar, G.N. and Bax, A. (1993) J. Magn. Reson., Ser. B, 102, 332–336.
Renault, J., Verchere-Beaur, C., Morgenstern-Badarau, I. and Piccioli, M. (1997) FEBS Lett., 401, 15–19.
Salgueiro, C., Turner, D. and Xavier, A. (1997) Eur. J. Biochem., 244, 721–734.
Satterlee, J. and La Mar, G.N. (1975) J. Am. Chem. Soc., 98, 2804–2808.
Sette, M., Paci, M., Desideri, A. and Rotilio, G. (1993) Eur. J. Biochem. 213, 391–397.
Sklenar, V., Miyashiro, H., Zon, G., Miles, H. and Bax, A. (1986) FEBS Lett., 208, 94–98.
Tu, K. and Gochin, M., in preparation.
Vila, A. and Fernandez, C. (1996) J. Am. Chem. Soc. 118, 7291–7998.
Wimperes, S. and Bodenhausen, G. (1989) Mol. Phys., 66, 897.
Wu, F. and Kurtz, D. (1989) J. Am. Chem. Soc., 111, 6563–6572.
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Gochin, M. Nuclear Magnetic Resonance Characterization of a Paramagnetic DNA-drug Complex with High Spin Cobalt; Assignment of the 1H and 31P NMR Spectra, and Determination of Electronic, Spectroscopic and Molecular Properties. J Biomol NMR 12, 243–257 (1998). https://doi.org/10.1023/A:1008289724077
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DOI: https://doi.org/10.1023/A:1008289724077