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. 2019 Nov 7;10:2572. doi: 10.3389/fmicb.2019.02572

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Copyright © 2019 Creighbaum, Ticak, Shinde, Wang and Ferguson.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Reconstitution of GB:CoM activity in vitro with purified recombinant proteins. Loss of the free thiol group on CoM was monitored at 412 nm, using Ellman’s reagent. Addition of GB resulted in a significant decrease in the amount of HS-CoM that is detectable (blue diamond), indicating an intact GB:CoM methyl transfer pathway. Methylation of CoM was not detectable when TMA (red square) or choline (gray triangle) served as the methyl donors. The assays contained 10 μg of MV8460 and 5 μg each of the remaining proteins. Assays in which MV8460, MV8465, MV10335, or MV10360 were omitted lacked GB:CoM activity (data not shown). Error bars represent standard deviations (n = 3).