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Details and methods are provided in the supporting material on Science Online.
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The expression of the yfgB or cfr genes in an E. coli methionine auxotroph cultured in the presence of [methyl-d3]methionine results in RlmN and Cfr containing deuterated methionine residues. More importantly, any methylated residue would bear a deuterated methyl group, because methionine is used biosynthetically to make SAM, which donates the methyl group in vivo during production of the protein.
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The peak at m/z = 252.1 in the black traces (RlmN and Cfr reactions) derives predominantly from abortive cleavage of SAM (fig. S4), in which the 5′-dA• abstracts a hydrogen atom that is ultimately solvent-derived, but also from a fraction of the enzyme that is isolated in its unmethylated state and is remethylated by unlabeled SAM.
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37
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79955532232
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note
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Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
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