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. 2015 Mar 4;15:59. doi: 10.1186/s12866-015-0387-7

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© Mitchell et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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VieB is a dose-dependent inhibitor of phosphotransfer. (A) Purified GST-VieS-C was incubated with ³²P-ATP-γ either alone (lane 1) or in the presence of equimolar VieA-His₆ (lane 2) and 5 μM VieB (lane 3) for 30 minutes at 30°C. (B) GST-VieS-C was incubated with ³²P-ATP-γ in the presence of equimolar VieA-His₆ and either 0, 0.25, 0.5, 1, 2, 4, or 8 μM VieB for 30 minutes at 30°C. Samples were stopped with the addition of 2X-denaturing sample buffer and separated using a 10% SDS-Page gel. Proteins labeled with ³²P were observed by radioautography. Figure panels are from the same experiment and exposed for the same amount of time. The radioautographs shown are a representative of three replicates. The band just below GST-VieS-C is a VieS degradation band, which is present in all figures and should be ignored.