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

Sampo Mäntylahti; Helena Tossavainen; Maarit Hellman; Perttu Permi

doi:10.1007/s10858-009-9373-4

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

J Biomol NMR. 2009 Nov;45(3):301-10. doi: 10.1007/s10858-009-9373-4.

Authors

Sampo Mäntylahti¹, Helena Tossavainen, Maarit Hellman, Perttu Permi

Affiliation

¹ Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR Laboratory, University of Helsinki, P.O. Box 65, 00014 Helsinki, Finland.

PMID: 19768387
DOI: 10.1007/s10858-009-9373-4

Abstract

An improved pulse sequence, intraresidual i(HCA)CO(CA)NH, is described for establishing solely (13)C'(i), (15)N(i), (1)HN(i) connectivities in uniformly 15N/13C-labeled proteins. In comparison to the "out-and-back" style intra-HN(CA)CO experiment, the new pulse sequence offers at least two-fold higher experimental resolution in the (13)C' dimension and on average 1.6 times higher sensitivity especially for residues in alpha-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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbon Isotopes / chemistry*
Humans
Nitrogen Isotopes / chemistry*
Nuclear Magnetic Resonance, Biomolecular / methods*
Proteins / chemistry*
Ubiquitin / chemistry

Substances

Carbon Isotopes
Nitrogen Isotopes
Proteins
Ubiquitin