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Calcium-Binding Protein Protocols Calcium-Binding Protein Protocols

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Shape and Dynamics of <strong>Calcium</strong>-<strong>Binding</strong> <strong>Protein</strong>s 297<br />

11. As can be seen from Eq. 7, larger experimental errors yield smaller χ 2 values,<br />

leading to the acceptance of simpler models. They are also propagated to produce<br />

larger uncertainties of the estimated parameters. This in turn diminishes the significance<br />

levels for the acceptance of models of increased complexity. Whereas<br />

high signal-to-noise spectra will produce desirably small experimental errors,<br />

uncontrollable sources of noise may limit the attainable uncertainties (see also<br />

Note 7).<br />

12. In the absence of structural information, the magnitude of the diffusion tensor<br />

may be estimated from the distribution of T 1/T 2 ratios (51).<br />

13. Some authors have suggested the use of entirely dynamics-based selection<br />

criteria (2).<br />

Acknowledgments<br />

J. M. Werner and I. D. Campbell thank the Wellcome Trust and the OCMS<br />

for support. The Oxford Centre for Molecular Sciences is funded by MRC,<br />

BBSRC, and EPSRC. A. K. Downing is a Wellcome Trust Senior Research<br />

Fellow, and she also thanks the support of the BBSRC and the MRC.<br />

References<br />

1. Kay, L. E. (1998) <strong>Protein</strong> dynamics from NMR. Nat. Struct. Biol. 5, 513–517.<br />

2. Barbato, G., Ikura, M., Kay, L. E., Pastor, R. W., and Bax, A. (1992) Backbone<br />

dynamics of calmodulin studied by N–15 relaxation using inverse detected<br />

2-dimensional NMR-spectroscopy — the central helix is flexible. Biochemistry 31,<br />

5269–5278.<br />

3. Baldellon, C., Alattia, J. R., Strub, M. P., Pauls, T., Berchtold, M. W., Cave, A., and<br />

Padilla, A. (1998) N–15 NMR relaxation studies of calcium-loaded parvalbumin<br />

show tight dynamics compared to those of other EF-hand proteins. Biochemistry<br />

37, 9964–9975.<br />

4. Paakkonen, K., Annila, A., Sorsa, T., Pollesello, P., Tilgmann, C., Kilpelainen, I.,<br />

et al. (1998) Solution structure and main chain dynamics of the regulatory domain<br />

(residues 1–91) of human cardiac troponin C. J. Biol. Chem. 273, 15,633–15,638.<br />

5. Gagne, S. M., Tsuda, S., Spyracopoulos, L., Kay, L. E., and Sykes, B. D. (1998)<br />

Backbone and methyl dynamics of the regulatory domain of troponin C: anisotropic<br />

rotational diffusion and contribution of conformational entropy to calcium<br />

affinity. J. Mol. Biol. 278, 667–686.<br />

6. Kordel, J., Skelton, N. J., Akke, M., Palmer, A. G., and Chazin, W. J. (1992) Backbone<br />

dynamics of calcium-loaded calbindin-D(9k) studied by 2-dimensional proton-detected<br />

N–15 NMR-spectroscopy. Biochemistry 31, 4856–4866.<br />

7. Werner, J. M., Knott, V., Handford, P. A., Campbell, I. D., and Downing, A. K.<br />

(2000) Backbone dynamics of a cbEGF domain pair in the presence of calcium.<br />

J. Mol. Biol. 296, 1065–1078.<br />

8. Dietz, H. C. and Pyeritz, R. E. (1995) Mutations in the human gene for fibrillin-1<br />

(FBN1) in the Marfan syndrome and related disorders. Hum. Mol. Genet. 4,<br />

1799–1809.

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