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. 2018 Jun 30;46(17):9160–9169. doi: 10.1093/nar/gky592

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — In vitro mo⁵U forming activity of TrmR. (A) On the left, HPLC analysis of TrmR directed the formation of mo⁵U (solid line) versus control which contains no TrmR (dashed line). The peak corresponding mo⁵U eluted from the C-18 column at 8.6 min which has been confirmed by spiking with an authentic compound (right panel). (B) The assay sample was analyzed by LC-MS/MS in positive mode, where the peak corresponding to mo⁵U ([C₁₀H₁₅N₂O₇]⁺, calculated m/z = 275.088) was selected (left) and the subsequent fragmentation by ESI was confirmed as detection of 5-methoxyuracil ([C₅H₇N₂O₃]⁺, calculated m/z = 143.046) in MS/MS (right).