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Simultaneous detection and deconvolution of congested NMR spectra containing three isotopically labeled species.

ABSTRACT: We present a procedure for isolating subspectra corresponding to individual protein or peptide components in a ternary mixture or complex. Each of the three-component species is labeled differently: species A uniformly with 15N, species B uniformly with 15N and 13C, and species C uniformly with 15N but selectively with 13C' or 13Calpha. By using the dual carbon label selective HSQC (DCLS-HSQC) pulse sequence and exploiting differences in 1J 15N-13C coupling patterns to filter selected 15N resonances from detection during a constant time period, a subspectrum from each species can be generated from three spectra acquired from a single sample. Many important biological pathways involve dynamic interactions among members of multicomponent protein assemblies, and this approach offers a powerful way to monitor such processes.

SUBMITTER: Masterson LR

PROVIDER: S-EPMC2662704 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature

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Simultaneous detection and deconvolution of congested NMR spectra containing three isotopically labeled species.

Masterson Larry R LR Tonelli Marco M Markley John L JL Veglia Gianluigi G

Journal of the American Chemical Society 20080531 25

We present a procedure for isolating subspectra corresponding to individual protein or peptide components in a ternary mixture or complex. Each of the three-component species is labeled differently: species A uniformly with 15N, species B uniformly with 15N and 13C, and species C uniformly with 15N but selectively with 13C' or 13Calpha. By using the dual carbon label selective HSQC (DCLS-HSQC) pulse sequence and exploiting differences in 1J 15N-13C coupling patterns to filter selected 15N resona ...[more]

PMID: 18512910

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