Helena, Kovacs and Gossert, Alvar (2014) Improved NMR experiments with 13C-isotropic mixing for assignment of aromatic and aliphatic side chains in labelled proteins. J Biomol NMR, 58 (2). pp. 101-112. ISSN 1573-5001

Abstract

Three improved 13C-13C-spinlock experiments for side chain assignments of isotope labelled proteins in liquid state are presented. These are based on wide bandwidth spinlock techniques that have become possible with contemporary cryogenic probes. The first application, the H(CaliCaro)H-TOCSY, is an HCCH-TOCSY in which all CHn moieties of a protein are detected in a single experiment, including the aromatic ones. This enables unambiguous assignment of aromatic amino acids in a single, highly sensitive experiment. In the second application, the 13C-detected Call-TOCSY, magnetization transfer comprises all carbons – aliphatic, aromatic as well as the carbonyl carbons – making the complete carbon assignment possible using one spectrum only. Thirdly, in the frequently used HC(CCO)NH experiment the long C-carbonyl refocused INEPT step is replaced by direct 13C-13C-TOCSY magnetization transfer from side chain carbons to the backbone carbonyls. The resulting HC(CCO)NH experiment minimizes relaxation losses because it is shorter and represents a more sensitive alternative particularly for larger proteins.

Item Type:	Article
Additional Information:	Same content as Conference Presentation submitted to OAK on same day.
Date Deposited:	14 Jun 2016 23:45
Last Modified:	04 Jul 2016 23:46
URI:	https://oak.novartis.com/id/eprint/9784