A novel protocol based on HN(C)N for rapid resonance assignment in (¹⁵N, ¹³C) labeled proteins: implications to structural genomics

Abstract

A novel protocol, based on the HN(C)N experiment, has been developed for rapid assignment of backbone H^N and ¹⁵N resonances in (¹⁵N, ¹³C) labeled proteins. The protocol exploits the directly observable ¹⁵N and H^N sequential correlations and the distinctive peak patterns in the different planes of the HN(C)N spectrum, depending upon the nature of the residues displaying the correlations. Glycines and prolines, which are responsible for the distinctive features, provide many check/start points for the sequential walks. These features enhance the speed of data analysis and render side chain assignments less crucial for the success of the assignments. The application of the protocol has been demonstrated with FK506 binding protein (FKBP, molecular mass 12 kDa).