Abstract
We introduce a solid-state NMR technique for selective detection of a residue pair in multiply labeled proteins to obtain site-specific structural constraints. The method exploits the frequency-offset dependence of cross polarization to achieve 13COi → 15Ni → 13Cαi transfer between two residues. A 13C, 15N-labeled elastin mimetic protein (VPGVG)n is used to demonstrate the method. The technique selected the Gly3 Cα signal while suppressing the Gly5 Cα signal, and allowed the measurement of the Gly3 Cα chemical shift anisotropy to derive information on the protein conformation. This residue-pair selection technique should simplify the study of protein structure at specific residues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baldus, M., Petkova, A.T., Herzfeld, J. and Griffin, R.G. (1998) Mol. Phys., 95, 1197–1207.
Cook, W.J., Einspahr, H., Trapane, T.L., Urry, D.W. and Bugg, C.E. (1980) J. Am. Chem. Soc., 1102, 5502–5505.
Egorova-Zachernyuk, T.A., Hollander, J., Fraser, N., Gast, P., Hoff, A.J., Cogdell, R., Groot, H.J.d. and Baldus, M. (2001) J. Biomol. NMR, 19, 243–253.
Havlin, R.H., Le, H., Laws, D.D., deDios, A.C. and Oldfield, E. (1997) J. Am. Chem. Soc., 119, 11951–11958.
Hong, M. (1999) J. Magn. Reson., 139, 389–401.
Hong, M. (2000) J. Am. Chem. Soc., 122, 3762–3770.
Hong, M. and Jakes, K. (1999) J. Biomol. NMR, 14, 71–74.
Huang, L., McMillan, R.A., Apkarian, R.P., Pourdeyhimi, B., Conticello, V.P. and Chaikof, E.L. (2000) Macromolecules, 33, 2989–2997.
Jaroniec, C.P., Tounge, B.A., Herzfeld, J. and Griffin, R.G. (2001) J. Am. Chem. Soc., 123, 3507–3519.
Kikuchi, J. and Asakura, T. (1999) J. Biochem. Biophys. Meth., 38, 203–208.
Li, B., Alonso, O.V. and Daggett, V. (2001) J. Mol. Biol., 305, 581–592.
Liu, S.F., Mao, J.D. and Schmidt-Rohr, K. (2001) J. Magn. Reson., in press.
McMillian, R.A. and Conticello, V.P. (2000) Macromolecules, 33, 4809–4821.
McMillian, R.A., Lee, T.A.T. and Conticello, V.P. (1999) Macromolecules, 32, 3643–3648.
Nomura, K., Takegoshi, K., Terao, T., Uchida, K. and Kainosho, M. (2000) J. Biomol. NMR, 17, 111–123.
Pines, A., Gibby, M.G. and Waugh, J.S. (1973) J. Chem. Phys., 59, 569–590.
Rienstra, C.M., Hohwy, M., Hong, M. and Griffin, R.G. (2000) J. Am. Chem. Soc., 122, 10979–10990.
States, D.J., Haberkorn, R.A. and Ruben, D.J. (1982) J. Magn. Reson., 48, 286–292.
Takegoshi, K., Nomura, K. and Terao, T. (1995) Chem. Phys. Lett., 232, 424–428.
Takegoshi, K., Nomura, K. and Terao, T. (1997) J. Magn. Reson., 127, 206–216.
Tjandra, N. and Bax, A. (1997) J. Am. Chem. Soc., 119, 9576–9577.
Tycko, R., Dabbagh, G. and Mirau, P. (1989) J. Magn. Reson., 85, 265–274.
Urry, D.W. (1988) J. Prot. Chem., 7, 1–34.
Urry, D.W. (1997) In On the Molecular Structure, Function and Pathology of Elastin: The Gotte Stepping Stone, Tamburro, A.M. (Ed.), Armento, Potenza, Italy, pp. 11–22.
Urry, D.W. (1999) Trends Biotechnol., 7, 249–257.
Urry, D.W., Chang, D.K., Krishna, N.R., Huang, D.H., Trapane, T.L. and Prasad, K.U. (1989) Biopolymers, 28, 819–833.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hong, M., McMillan, R.A. & Conticello, V.P. Measurement of conformational constraints in an elastin-mimetic protein by residue-pair selected solid-state NMR. J Biomol NMR 22, 175–179 (2002). https://doi.org/10.1023/A:1014291305148
Issue Date:
DOI: https://doi.org/10.1023/A:1014291305148