Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2015 Aug 31;43(22):10713–10721. doi: 10.1093/nar/gkv870

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2015. 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 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

PMC Copyright notice

Figure 3. — Oxidation activities of A212 and V293 mutants. (A) LC–MS/MS quantification of 5mC and its oxidized derivatives after a 10-min reaction of NgTet1 WT and variant proteins. Inserted is a SDS-PAGE gel (bottom) of the proteins (WT and 8 mutants) used for activity (see Supplemental Figure S3a). The protein concentrations were adjusted and equal amount of enzymes (WT and mutants) were used in each reaction. Error bars indicate standard error (s.e.) of the mean value from three independent experiments. (B and C) The time courses of quantitative LC–MS/MS measurements of 5mC (black) disappearance and formation of 5hmC (blue), 5fC (magenta) and 5caC (green) by WT (panel b) or 5hmC by A212V (panel c). (D) A model of V212 with two alternative conformations (in green or grey). The conformation in grey would clash with 5hmC (indicated by a star). (E) A model of L212. Only one of the three possible conformations was shown (in yellow), which would clash with Y141. The other two conformations would clash with V293 or F295 (in the background away from the reader). (F) A model of N212 (magenta) superimposed with L212 (yellow). The planar side chain conformation allowed N212 to be accommodated near Y141. (G) Comparison of V212, L212 and N212. (H and I) LC–MS/MS quantification of oxidation of a 5hmC- (panel G) or 5fC-containing oligo (panel h) after 10-min reactions of NgTet1 WT and mutant variants. Error bars indicate standard error (s.e.) of the mean value from three independent experiments.