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An intraresidual i(HCA)CO(CA)NH experiment for the assignment of main-chain resonances in 15N, 13C labeled proteins


Abstract An improved pulse sequence, intraresidual i(HCA)CO(CA)NH, is described for establishing solely 13Câ?²(i), 15N(i), 1HN(i) connectivities in uniformly 15N/13C-labeled proteins. In comparison to the â??out-and-backâ?? style intra-HN(CA)CO experiment, the new pulse sequence offers at least two-fold higher experimental resolution in the 13Câ?² dimension and on average 1.6 times higher sensitivity especially for residues in α-helices. Performance of the new experiment was tested on a small globular protein ubiquitin and an intrinsically unfolded 110-residue cancer/testis antigen CT16/PAGE5. Use of intraresidual i(HCA)CO(CA)NH experiment in combination with the established HNCO experiment was crucial for the assignment of highly disordered CT16.

• Content Type Journal Article
• Pages 301-310
• DOI 10.1007/s10858-009-9373-4
• Authors
  • Sampo Mäntylahti, University of Helsinki Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory P.O. Box 65 00014 Helsinki Finland
  • Helena Tossavainen, University of Helsinki Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory P.O. Box 65 00014 Helsinki Finland
  • Maarit Hellman, University of Helsinki Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory P.O. Box 65 00014 Helsinki Finland
  • Perttu Permi, University of Helsinki Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory P.O. Box 65 00014 Helsinki Finland

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738
  • Journal Volume Volume 45
  • Journal Issue Volume 45, Number 3

Source: Journal of Biomolecular NMR