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. 2018 Jul 26;9:1688. doi: 10.3389/fmicb.2018.01688

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Copyright © 2018 Ongey, Giessmann, Fons, Rappsilber, Adrian and Neubauer.

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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Expression test and MS²-ESI-Orbitrap spectrum of gel-extracted His6-preRumA. (A) SDS-PAGE analysis of purified soluble (lane 1) and insoluble (lane 2) His6-preRumA produced in the absence of RumM (i.e., extracts from strain WLEOrA). (B) Analysis of purified control extracts from strain WLEOrA (Lane 1) and extracts from strain WLEOrA/M, showing soluble production of His6-RumM (Lane 2). (C) Soluble His6-preRumA extracted from strain WLEOrA/M (Lanes 1 and 2). (D) Complete fragmentation of His6-preRumA, following tryptic digestion. None of the precursor ions resulting from the LC-MS analysis of the tryptic digest indicated fragmentation restrictions at the positions where thioether cross-bridges were expected to be formed. Consequently, there was no ring structure in the product measured. cm, carbamidomethylation; ox, oxidation. Full scans of sliced gel pieces in A, B and C are presented in Supplementary Figure S9.