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Improved labeling strategy for 13C relaxation measurements of methyl groups in proteins

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Abstract

Selective incorporation of 13C into the methyl groupsof protein side chains is described as a means for simplifying themeasurement and interpretation of 13C relaxation parameters.High incorporation (>90%) is accomplished by using pyruvate(3-13C, 99%) as the sole carbon source in the growthmedia for protein overexpression in E. coli. This improved labeling schemeincreases the sensitivity of the relaxation experiments by approximatelyfivefold when compared to randomly fractionally 13C-labeledprotein, allowing high-quality measurements on relatively dilute (<1 mM)protein samples at a relatively low cost.

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Authors and Affiliations

  1. Departments of Biological Sciences, Biophysical Sciences and Chemistry, and Center for Structural Biology, State University of New York at Buffalo, 816 Natural Sciences and Mathematics Complex, Buffalo, NY, 14260-3000, U.S.A

    Andrew L. Lee, Jeffrey L. Urbauer & A. Joshua Wand

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  1. Andrew L. Lee
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  2. Jeffrey L. Urbauer
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  3. A. Joshua Wand
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Lee, A.L., Urbauer, J.L. & Wand, A.J. Improved labeling strategy for 13C relaxation measurements of methyl groups in proteins. J Biomol NMR 9, 437–440 (1997). https://doi.org/10.1023/A:1018311013338

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