One-sample approach to determine the relative orientations of proteins in ternary and binary complexes from residual dipolar coupling measurements

Marco Tonelli, Larry R. Masterson, Gabriel Cornilescu, John L. Markley, Gianluigi Veglia

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

(Figure Presented) We present a procedure that supports the acquisition of 1H-15N residual dipolar coupling (RDC) values for individual subunits in binary or ternary protein assemblies from a single experimental sample. Our method relies on asymmetric labeling of each subunit with the following scheme: species A uniformly with 15N, species B uniformly with 15N and 13C, and species C uniformly with 15N but selectively with 13C′ or 13C α. Because only a single sample is required, the approach obviates the need for preparing multiple samples and eliminates potential errors introduced from differences in sample conditions. Because numerous biological processes rely on protein assemblies or transient interactions, this method should be well suited for a wide range of future applications.

Original languageEnglish (US)
Pages (from-to)14138-14139
Number of pages2
JournalJournal of the American Chemical Society
Volume131
Issue number40
DOIs
StatePublished - Oct 14 2009

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