Abstract
Defining the self-association state of a molecule in solution can be an important step in NMR-based structure determination. This is particularly true of peptides, where there can be a relatively small number of long-range interactions and misinterpretation of an intermolecular NOE as an intramolecular contact can have a dramatic influence on the final calculated structure. In this paper, we have investigated the use of translational self-diffusion coefficient measurements to detect self-association in aqueous trifluoroethanol of three peptides which are analogues of the C-terminal region of human neuropeptide Y. Experimentally measured diffusion coefficients were extrapolated to D0, the limiting value as the peptide concentration approaches zero, and then converted to D20,w, the diffusion coefficient after correction for temperature and the viscosity of the solvent. A decrease in D20,w of about 16% was found for all three peptides in aqueous TFE (30% by volume) compared with water, which is in reasonable agreement with the expected decrease upon dimerisation, the presence of which was indicated by sedimentation equilibrium measurements. Apparent molecular masses of these peptides in both solutions were also calculated from their diffusion coefficients and similar results were obtained. Several potential internal standards, including acetone, acetonitrile, dimethylsulfoxide and dioxane, were assessed as monitors of solution viscosity over a range of trifluoroethanol concentrations. Compared with independent measurements of viscosity, acetonitrile was the most accurate standard among these four. The practical limitations of a quantitative assessment of peptide self-association from translational diffusion coefficients measured by PFGNMR, including the calculation of apparent molecular mass, are also discussed.
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Altieri, A.S., Hilton, D.P. and Byrd, R.A. (1995) J. Am. Chem. Soc., 117, 7566–7567.
Barden, J.A. (1995) Biochem. Biophys. Res. Commun., 215, 264–271.
Barden, J.A., Cuthbertson, R.M., Potter, E.K., Selbie, L.A. and Tseng, A. (1994) Biochim. Biophys. Acta, 1206, 191–196.
Barnham, K.J., Catalfamo, F., Pallaghy, P.K., Howlett, G.J. and Norton, R.S. (1999) Biochim. Biophys. Acta, 1435, 127–137.
Callaghan, P.T., Gros, M.A.L. and Pinder, D.N. (1983) J. Chem. Phys., 79, 6372–6381.
Cantor, C.R. and Schimmel, P.R. (1980) Biophysical Chemistry, Part II: Techniques for the Study of Biological Structure and Function, W.H. Freeman, New York, NY, pp. 539–590.
Chen, A., Johnson, C.S. Jr., Lin, M. and Shapiro, M.J. (1998) J. Am. Chem. Soc., 120, 9094–9095.
Chen, A., Wu, D. and Johnson, C.S. Jr.(1995) J. Phys. Chem., 99, 828–834.
Cowley, D.J., Hoflack, J.M., Pelton, J.T. and Saudek, V. (1992) Eur. J. Biochem., 205, 1099–1106.
Dingley, A.J., Mackay, J.P., Chapman, B.E., Morris, M.B., Kuchel, P.W., Hambly, B.D. and King, G.F. (1995) J. Biomol. NMR, 6, 321–328.
Doran, S.J. and Décorps, M. (1995) J. Magn. Reson., A 117, 311–316.
Gibbs, S.J. and Johnson, C.S. Jr. (1991) J. Magn. Reson., 93, 395–402.
Grundemar, L. (1997) In Neuropeptide Y and Drug Development (Grundemar, L. and Bloom, S.R, Eds.), Academic Press, San Diego, CA, pp. 1–11.
Grundemar, L. and Hå kanson, R. (1994) Trends Pharm. Sci., 15, 153–158.
Houston, M.E., Gannon, C.L., Kay, C.M. and Hodges, R.S. (1995) J. Pept. Sci., 1, 274–282.
Jones, J.A., Wilkins, D.K., Smith, L.J. and Dobson, C.M. (1997) J. Biomol. NMR, 10, 199–203.
Kirby, D.A., Britton, K.T., Aubert, M.L. and Rivier, J.E. (1997) J. Med. Chem., 40, 210–215.
Kuntz, I.D.Jr.and Kauzmann, W. (1974) In Advances in Protein Chemistry, Vol. 28 (Anfinsen, C.B., Edsall, J.T. and Richards, F.M., Eds.), Academic Press, New York, NY, pp. 239–345.
Lapham, J., Rife, J.P., Moore, P.B. and Crothers, D.M. (1997) J. Biomol. NMR, 10, 255–262.
Lin, M., Shapiro, M.J. and Wareing, J.R. (1997) J. Am. Chem. Soc., 119, 5249–5250.
Longsworth, L.G. (1960) J. Phys. Chem., 64, 1914–1917.
MacPhee, C.E., Perugrini, M.A., Sawyer, W.H. and Howlett, G.J. (1997) FEBS Lett., 416, 265–268.
Monks, S.A., Karagianis, G., Howlett, G.J. and Norton, R.S. (1996) J. Biomol. NMR, 8, 379–390.
Mulhern, T.D., Howlett, G.J., Reid, G.E., Simpson, R.J., McColl, D.J., Anders, R.F. and Norton, R.S. (1995) Biochemistry, 34, 3479–3491.
Natarajan, G. (1989) Data Book on the Viscosity of Liquids, Hemisphere, New York, NY.
Nelson, J.W. and Kallenbach, N.R. (1986) Proteins Struct. Funct. Genet., 1, 211–217.
Nordmann, A., Blommers, M.J.J., Fretz, H., Arvinte, T. and Drake, A.F. (1999) Eur. J. Biochem., 261, 216–226.
Pan, H., Barany, G. and Woodward, C. (1997) Protein Sci., 6, 1985–1992.
Perkins, S.J. (1986) Eur. J. Biochem., 157, 169–180.
Potter, E.K., Barden, J.A., McCloskey, M.J.D., Selbie, L.A., Tseng, A., Herzog, H. and Shine, J. (1994) Eur. J. Pharmacol., 267, 253–262.
Ralston, G. (1993) Introduction to Analytical Ultracentrifugation, Beckman Instruments, Inc., CA.
Rist, B., Zerbe, O., Ingenhoven, N., Scapozza, L., Peers, C., Vaughan, P.F.T., McDonald, R.L., Wieland, H.A. and Beck-Sickinger, A.G. (1996) FEBS Lett., 394, 169–173.
Schuck, P., MacPhee, C.E. and Howlett, G.J. (1998) Biophys. J., 74, 466–474.
Sönnichsen, F.D., Van Eyk, J.E., Hodges, R.S. and Sykes, B.D. (1992) Biochemistry, 31, 8790–8798.
Stejskal, E.O. and Tanner, J.E. (1965) J. Chem. Phys., 42, 288–292.
Teller, D.C., Swanson, E. and de HaËn, C. (1979) Methods Enzymol., 61, 103–124.
Weast, R.C. (1984) CRC Handbook of Chemistry and Physics, 64th ed., CRC Press, Boca Raton, FL.
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Yao, S., Howlett, G.J. & Norton, R.S. Peptide self-association in aqueous trifluoroethanol monitored by pulsed field gradient NMR diffusion measurements. J Biomol NMR 16, 109–119 (2000). https://doi.org/10.1023/A:1008382624724
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DOI: https://doi.org/10.1023/A:1008382624724