Summary
A 3D NMR technique is described which correlates the amide proton and nitrogen resonances of an amino acid residue with the Cα chemical shift of its preceding residue. The technique uses a relay mechanism, transferring magnetization from15N to13Cα via the intervening carbonyl nucleus. This method for obtaining sequential connectivity is less sensitive to large line widths than the alternative HNCA experiment. The technique is demonstrated for the protein calmodulin, complexed with a 26 amino acid fragment of skeletal muscle myosin light chain kinase.
This is a preview of subscription content, log in via an institution to check access.
Abbreviations
- CaM:
-
Calmodulin
- HCACO:
-
α-proton to α-carbon to carbonyl correlation
- H(CA)NHN:
-
α-proton (via α-carbon) to nitrogen to amide proton correlation
- HMQC:
-
heteronuclear multiple quantum correlation
- HNCA:
-
amide proton to nitrogen to C α-carbon correlation
- M13:
-
a 26-residue fragment of the CaM-binding domain of skeletal muscle myosin light chain kinase comprising residues 577–602.
References
Bax, A., Griffey, R.H. and Hawkins, B.L. (1983)J. Magn. Reson.,55, 301–315.
Bendall, M.R., Pegg, D.T. and Doddrell, D.M. (1983)J. Magn. Reson.,52, 81–117.
Burum, D.P. and Ernst, R.R. (1990)J. Magn. Reson. 39, 533–538.
Ernst, R.R., Bodenhausen, G. and Wokaun, A. (1987)Principles of Nuclear Magnetic Resonance in One and Two Dimensions, Clarendon Press, Oxford, pp. 25–32.
Ikura, M., Kay, L.E. and Bax, A. (1990)Biochemistry,29, 4659–4667.
Ikura, M., Kay, L.E., Krinks, M. and Bax, A. (1991)Biochemistry, in press.
Kay, L.E., Marion, D. and Bax, A. (1989)J. Magn. Reson.,84, 72–84.
Kay, L.E., Ikura, M., Tschudin, R. and Bax, A. (1990)J. Magn. Reson. 89, 496–514.
Kay, L.E., Ikura, M. and Bax, A. (1991)J. Magn. Reson.,91, 84–92.
Llinas, M., Wilson, D. and Klein, M.P. (1977)J. Am. Chem. Soc.,99, 6846–6849.
London, R.E. (1990)J. Magn. Reson.,86, 410–415.
Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989a)J. Magn. Reson.,85, 393–399.
Marion, D., Ikura, M. and Bax, A. (1989b)J. Magn. Reson.,85, 425–430.
Morris, G.A. and Freeman, R. (1979)J. Am. Chem. Soc.,99, 760–761.
Niemczura, W.P., Helms, G.L., Chesnick, A.S., Moore, R.E. and Bornemann, V. (1989)J. Magn. Reson.,81, 635–640.
Oh, B.H., Westler, W.M., Derba, P. and Markley, J.L. (1988)Science Washington DC,240, 908–911.
Spera, S., Ikura, M. and Bax, A. (1991)J. Biomolec. NMR, in press.
Stockman, B.J., Reilly, M.D., Westler, W.M., Ulrich, E.L. and Markley, J.L. (1989)Biochemistry,28, 230–236.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bax, A., Ikura, M. An efficient 3D NMR technique for correlating the proton and15N backbone amide resonances with the α-carbon of the preceding residue in uniformly15N/13C enriched proteins. J Biomol NMR 1, 99–104 (1991). https://doi.org/10.1007/BF01874573
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01874573