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. 2019 Dec 23;117(1):629–634. doi: 10.1073/pnas.1906748117

Fig. 2.

Fig. 2.

Nucleotide-dependent dissociation of 70S ribosomes by Ms-HflX in vitro. (AF) Dissociation of 70S ribosomes (0.2 μM) was carried out in the presence of 3.0 μM Ms-HflX6his in HMA-7 buffer in the presence of 1 mM GTP, ATP, or GMP-PNP at 37 °C for 45 min and examined using a 5-mL analytical 10% to 40% SDGC. Reactions lacking either nucleotide or HflX were included as controls. Percentage area under the curve was calculated for 70S and 50S peaks, using PeakChart (v. 2.08, Brandel), and expressed as a ratio of 70S:50S. Data represent mean ± SD, n = 3. (G) The samples were collected using the Brandel Teledyne ISCO gradient fractionation system, methanol-chloroform precipitated, followed by immunoblotting with anti-his antibody to determine the presence of Ms-HflX6his in each fraction.