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. 2013 Jan 15;41(5):3217–3227. doi: 10.1093/nar/gks1475

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — In vitro binding assay for RbgA mutants to 50S subunit under different nucleotide conditions. Purified RbgA-his₆ or RbgA mutant-his₆ (50 nM) was incubated with purified 50S (10 nM) subunit and GDP, GTP or GMP-PNP (400 µM) at 37°C. The presence of RbgA was tested by western blot with custom RbgA-antibody and quantified. Percentages were generated by assigning RbgA + 50S + GMP-PNP as 100%. Lane 1: RbgA + 50S + GDP; lane 2: RbgA + 50S + GTP; lane 3: RbgA + 50S + GMP-PNP; lane 4: A206D + 50S + GDP; lane 5: A206D + 50S + GTP; lane 6: A206D + 50S + GMP-PNP; lane 7: Y225A + 50S + GDP; lane 8: Y225A + 50S + GTP; lane 9: Y225A + 50S + GMP-PNP.