A sequential HNCA NMR pulse sequence for protein backbone assignment

Journal of Magnetic Resonance
A Meissner, O W Sørensen

Abstract

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(alpha)) 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(alpha) correlation.

Citations

Jul 21, 2010·Journal of Magnetic Resonance·Dinesh KumarRamakrishna V Hosur
Jan 15, 2005·Journal of Magnetic Resonance·Wolfgang BermelPaul R Vasos
May 18, 2010·Protein Expression and Purification·Filippo PrischiAnnalisa Pastore
Jun 26, 2002·Journal of Magnetic Resonance·Bernhard Brutscher
Apr 28, 2006·Journal of the American Chemical Society·Dominique P FruehGerhard Wagner
Sep 13, 2002·Journal of the American Chemical Society·Daniel NietlispachErnest D Laue

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