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Use of a copper-chelated lipid speeds up NMR measurements from membrane proteins.


ABSTRACT: Recent studies have demonstrated the abilities of solid-state NMR techniques to solve atomic-level-resolution structures and dynamics of membrane-associated proteins and peptides. However, high-throughput applications of solid-state NMR spectroscopy are hampered by long acquisition times due to the low sensitivity of the technique. In this study, we demonstrate the use of a paramagnetic copper-chelated lipid to enhance the spin-lattice relaxation and thereby speed up solid-state NMR measurements. Fluid lamellar-phase bicelles composed of a lipid, detergent, and the copper-chelated lipid and containing a uniformly (15)N-labeled antimicrobial peptide, subtilosin A, were used at room temperature. The use of a chelating lipid reduces the concentration of free copper and limits RF-induced heating, a major problem for fluid samples. Our results demonstrate a 6.2-fold speed increase and a 2.7-fold improvement in signal-to-noise ratio for solid-state NMR experiments under magic-angle spinning and static conditions, respectively. Furthermore, solid-state NMR measurements are shown to be feasible even for nanomole concentrations of a membrane-associated peptide.

SUBMITTER: Yamamoto K 

PROVIDER: S-EPMC2874097 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature

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Use of a copper-chelated lipid speeds up NMR measurements from membrane proteins.

Yamamoto Kazutoshi K   Xu Jiadi J   Kawulka Karen E KE   Vederas John C JC   Ramamoorthy Ayyalusamy A  

Journal of the American Chemical Society 20100501 20


Recent studies have demonstrated the abilities of solid-state NMR techniques to solve atomic-level-resolution structures and dynamics of membrane-associated proteins and peptides. However, high-throughput applications of solid-state NMR spectroscopy are hampered by long acquisition times due to the low sensitivity of the technique. In this study, we demonstrate the use of a paramagnetic copper-chelated lipid to enhance the spin-lattice relaxation and thereby speed up solid-state NMR measurements  ...[more]

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