Journal of Biomolecular NMR

, Volume 62, Issue 3, pp 281–290

Assignment of methyl NMR resonances of a 52 kDa protein with residue-specific 4D correlation maps

Article

Abstract

Methyl groups have become key probes for structural and functional studies by nuclear magnetic resonance. However, their NMR signals cluster in a small spectral region and assigning their resonances can be a tedious process. Here, we present a method that facilitates assignment of methyl resonances from assigned amide groups. Calculating the covariance between sensitive methyl and amide 3D spectra, each providing correlations to Cα and Cβ separately, produces 4D correlation maps directly correlating methyl groups to amide groups. Optimal correlation maps are obtained by extracting residue-specific regions, applying derivative to the dimensions subject to covariance, and multiplying 4D maps stemming from different 3D spectra. The latter procedure rescues weak signals that may be missed in traditional assignment procedures. Using these covariance correlation maps, nearly all assigned isoleucine, leucine, and valine amide resonances of a 52 kDa nonribosomal peptide synthetase cyclization domain were paired with their corresponding methyl groups.

Keywords

NMR resonance assignment Large proteins Methyl groups Nonribosomal peptide synthetases 

Supplementary material

10858_2015_9943_MOESM1_ESM.pdf (774 kb)
Supplementary material 1 (PDF 775 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  1. 1.Department of Biophysics and Biophysical ChemistryJohns Hopkins University School of MedicineBaltimoreUSA

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