Abstract
The mRNA cap-binding protein eIF4E is the limiting factor in the eIF4F translation initiation complex, which mediates the binding of the 40S ribosome to the mRNA. 15N relaxation studies have been used to characterize the backbone dynamics of deuterated eIF4E in a CHAPS micelle for the apoprotein, the m7GDP-bound form, and the dinucleotide (m7GpppA)-bound form, as well as for CHAPS-free eIF4E. Large differences in overall correlation time between the CHAPS-free form (11.8 ns) and samples containing different concentrations of CHAPS (15.9–19.4 ns) indicate that eIF4E is embedded in a large micelle in the presence of CHAPS, with a total molecular weight in the range of 40–60 kDa. CHAPS seems to restrict the mobility of the a2–b3 and a4–b5 loops which are thought to be embedded in the micelle. No significant changes in overall mobility were seen between the m7GDP-bound form, the m7GpppA-bound form, and the apoprotein. Amide hydrogen exchange data indicate the presence of slowly exchanging amides in two surface-exposed helices (a2 and a4), as well as the a4–b5 loop, indicating protection by the CHAPS micelle. The micelle covers the convex side of the protein away from the cap-binding site.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bartels, C., Xia, T.-H., Billeter, M., Güntert, P., and Wüthrich, K. (1995) J. Biomol. NMR, 6, 1–10.
Cavanagh, J., Palmer III, A.G., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 91, 429–436.
Cavanagh, J. and Rance, M. (1993) Annu Rep. NMR Spectrosc., 27, 1–58.
Chattopadhyay, A. and Harikumar, K.G. (1996) FEBS Lett., 391, 199–202.
Dayie, K.T., Wagner, G. and Lefèvre, J.F. (1996) Annu. Rev. Phys. Chem., 47, 243–282.
Edery, I., Altmann, M. and Sonenberg, N. (1988) Gene, 74, 517–525.
Farrow, N.A., Muhandiram, R., Singer, A.U., Pascal, S.M., Kay, C.M., Gish, G., Shoelson, S. E., Pawson, T., Forman-Kay, J.D., and Kay, L.E. (1994) Biochemistry, 33, 5984–6003.
Haghighat, A., Mader, S., Pause, A. and Sonenberg, N. (1995) EMBO J., 14, 5701–5709.
Ishima, R. and Nagayama, K. (1995a) Biochemistry, 34, 3162–3171.
Ishima, R. and Nagayama, K. (1995b) J. Magn. Reson., B108, 73–76.
Kay, L.E., Torchia, D.E. and Bax, A. (1989) Biochemistry, 28, 8972–8979.
Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663–10665.
Kraulis, P.J. (1991) J. Appl. Crystallogr., 24, 946–950.
Marcotrigiano, J., Gingras, A.-C., Sonenberg, N., and Burley, S.K. (1997) Cell, 89, 951–961.
Matsuo, H., Li, H., McGuire, A. M., Fletcher, C. M., Gingras, A.-C., Sonenberg, N. and Wagner G. (1997) Nat. Struct. Biol., 4, 717–724.
Merrick, W.C. and Hershey, J.W.B. (1996) In Translational Control (Eds., Hershey, J.W.B., Mathews, M.B. and Sonenberg, N.), Cold Spring Harbor Laboratory Press, Plainview, New York, NY, pp. 31–69.
Nicholls, A., Sharp, K.A. and Honig, B. (1991) Proteins, 11, 281–296.
Nirmala, N. R. and Wagner, G. (1988) J. Am. Chem. Soc., 110, 7557–7558.
Nirmala, N.R. and Wagner, G. (1989) J. Magn. Reson., 82, 659–661.
Palmer III, A.G., Cavanaugh J., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 93, 151–170.
Peng, J.W. and Wagner, G. (1992a) J. Magn. Reson., 98, 308–332.
Peng, J.W., and Wagner, G. (1992b) Biochemistry, 31, 8571–8586.
Peng, J.W. and Wagner, G. (1995) Biochemistry, 34, 16733–16752.
Schleucher, J., Schwendinger, M., Sattler, M., Schmidt, P., Schedletzky, O., Glaser, S.J., Sørenson, O.W., Griesinger, C. (1994) J. Biomol. NMR, 4, 301–306.
Wagner, G. (1997) Nat. Struct. Biol., 4, 841–844.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
McGuire, A.M., Matsuo, H. & Wagner, G. Internal and overall motions of the translation factor eIF4E: Cap binding and insertion in a CHAPS detergent micelle. J Biomol NMR 12, 73–88 (1998). https://doi.org/10.1023/A:1008214128792
Issue Date:
DOI: https://doi.org/10.1023/A:1008214128792