Abstract
Cyclase-associated proteins (CAPs) are highly conserved, ubiquitous actin binding proteins that are involved in microfilament reorganization. The N-termini of CAPs play a role in Ras signaling and bind adenylyl cyclase; the C-termini bind to G-actin. We report here the NMR characterization of the amino-terminal domain of CAP from Dictyostelium discoideum (CAP(1-226)). NMR data, including the steady state 1H-15N heteronuclear NOE experiments, indicate that the first 50 N-terminal residues are unstructured and that this highly flexible serine-rich fragment is followed by a stable, folded core starting at Ser 51. The NMR structure of the folded core is an α-helix bundle composed of six antiparallel helices, in a stark contrast to the recently determined CAP C-terminal domain structure, which is solely built by β-strands.
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References
Brünger, A.T. A System for X-Ray Crystallography and NMR, Yale University Press, New Haven, CT.
Cornilescu, G., Delaglio, F. and Bax, A. (1999) J. Biomol. NMR, 13, 289–302.
Farrow, N.A., Muhandiram, R., Singer, A.U., Pascal, S.M., Kay, C.M., Gish, G., Shoelson, S.E., Pawson, T., Foreman-Kay, J.D. and Kay, L.E. (1994) Biochemistry, 33, 5984–6003.
Field, J., Vojtek, A., Ballester, R., Bolger, G., Colicelli, J., Ferguson, K., Gerst, J., Kataoka, T., Powers, S., Riggs, M., Rodgers, L., Wieland, I., Wheland, B. and Wigler, M. (1990) Cell, 61, 319–327.
Gerst, J.E., Rodgers, L., Riggs, M. and Wigler, M. (1992) Proc. Natl. Acad. Sci. USA, 89, 4338–4342.
Goddard, T.D. and Kneller, D.G. (2000) 'SPARKY 3', University of California, San Francisco.
Gottwald, U., Brokamp, R., Karakesisoglou, I., Schleicher, M. and Noegel A.A. (1996) Mol. Biol. Cell, 7, 261–272.
Hubberstey, A.V. and Mottillo E.P. (2002) FASEB J., 16, 487–499.
Koradi, R., Billeter, M. and Wüthrich, K. (1996) J. Mol. Graph., 14, 52–55.
Ksiazek, D., Brandstetter, H., Israel, L., Bourenkov, G.P., Katchalova, G., Janssen, K.-P., Bartunik, H. D., Noegel, A.A., Schleicher, M. and Holak, T.A. (2003) Structure, 11, 1–20.
Mühlhahn, P., Bernhagen, J., Czisch, M., Georgescu, J., Renner, C., Ross, A., Bucala, R. and Holak, T.A. (1996) Protein Sci., 5, 2095–2103.
Nilges, M., Clore, G.M. and Gronenborn, A.M. (1988) FEBS Lett., 239, 129–136.
Nishida, Y., Shima, F., Sen, H., Tanaka, Y., Yanagihara, C., Yamawaki-Kataoka, Y., Kariya, K., and Kataoka, T. (1998) J. Biol. Chem., 273, 28019–28024.
Noegel, A.A., Rivero, F., Albrecht, R., Janssen, K.P., Koehler, J., Parent, C.A. and Schleicher, M. (1999) J. Cell Sci., 112, 3195–3203.
Powell, M. J. D. (1977) Math. Program., 12, 241–254.
Rehm, T., Mavoungou, Ch., Israel, L., Schleicher, M. and Holak, T.A. (2002a) J. Biomol. NMR, 23, 337–338.
Rehm, T., Huber, R. and Holak, T.A. (2002b) Structure, 10, 1613–1618.
Renner, C., Schleicher, M., Moroder, L. and Holak T.A. (2002) J. Biomol. NMR, 23, 23–33.
Senn, H., Eugster, A., Otting, G., Suter, F. and Wüthrich, K. (1987) Eur. Biophys. J., 14, 301–316.
Stevenson, V.A. and Theurkauf, W.E. (2000) Curr. Biol., 10, R695–697.
Weber, T., Baumgartner, R., Renner, C., Marahiel, M.A. and Holak, T.A. (2000) Structure, 8, 407–418.
Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY.
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Mavoungou, C., Israel, L., Rehm, T. et al. NMR structural characterization of the N-terminal domain of the adenylyl cyclase-associated protein (CAP) from Dictyostelium discoideum . J Biomol NMR 29, 73–84 (2004). https://doi.org/10.1023/B:JNMR.0000019513.86120.98
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DOI: https://doi.org/10.1023/B:JNMR.0000019513.86120.98