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Measurement of residual dipolar couplings in methyl groups via carbon detection

Authors
  • Williams, Robert V.
  • Yang, Jeong-Yeh
  • Moremen, Kelley W.
  • Amster, I. Jonathan
  • Prestegard, James H.
Type
Published Article
Journal
Journal of Biomolecular NMR
Publisher
Springer-Verlag
Publication Date
Apr 30, 2019
Volume
73
Issue
3-4
Pages
191–198
Identifiers
DOI: 10.1007/s10858-019-00245-5
Source
Springer Nature
Keywords
License
Yellow

Abstract

Residual dipolar couplings (RDCs) provide both structural and dynamical information useful in the characterization of biological macromolecules. While most data come from the interaction of simple pairs of directly bonded spin-1/2 nuclei (1H–15N, 1H–13C, 1H–1H), it is possible to acquire data from interactions among the multiple spins of 13C-labeled methyl groups (1H3–13C). This is especially important because of the advantages that observation of 13C-labeled methyl groups offers in working with very large molecules. Here we consider some of the options for measurement of methyl RDCs in large and often fully protonated proteins and arrive at a pulse sequence that exploits both J-modulation and direct detection of 13C. Its utility is illustrated by application to a fully protonated two domain fragment from the mammalian glycoprotein, Robo1, 13C-methyl-labeled in all valines.

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