A Sequential HNCA NMR Pulse Sequence for Protein Backbone Assignment

✍ Scribed by Axel Meissner; Ole Winneche Sørensen

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 90 KB
• Volume: 150
• Category: Article
• ISSN: 1090-7807
• DOI: 10.1006/jmre.2001.2317

✦ Synopsis

The conventional HNCA pulse sequence suffers from the ambiguity that it cannot distinguish inter-and intraresidue correlations because the one-bond and two-bond J(NC α ) coupling constants are of similar magnitude. This paper presents a novel pulse sequence, sequential HNCA, that leads to a spectrum exhibiting exclusively interresidue correlations. This important sequential information has so far usually been obtained by an HN(CO)CA experiment that for medium field strengths typically also is more sensitive than HNCA. However, for increasing static magnetic fields the chemical shift anisotropy relaxation mechanism of carbonyl carbons becomes more and more efficient, leading to a degradation of the HN(CO)CA sensitivity. Hence there is a point where the sequential HNCA experiment becomes the most sensitive option for sequential N-C α correlation.