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. 2021 Jan 15;49(4):1987–2004. doi: 10.1093/nar/gkaa1291

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — E. coli RecO is monomeric. (A) Sedimentation velocity c(s) distribution profiles of RecO (monitored at 230 nm) at 1.5, 4 and 8 μM in buffer BTP (pH 8.0) at 25°C. (B) The weight average sedimentation coefficient of RecO slightly decreases as a function of RecO concentration, expected for a pure, non-associating species (103–106). A linear extrapolation to zero RecO concentration yields s⁰ = 0.87 ± 0.04 S. From this we calculate = 2.32 ± 0.05 S, consistent with a RecO monomer. (C) Results of sedimentation equilibrium experiments (monitored at 230 nm) performed at 4 μM RecO at three rotor speeds 20,000 (blue), 25 000 (orange) and 30 000 (gray) rpm are shown in blue, orange and gray. Each equilibrium profile is described by a single exponential. The solid lines show the best global NLLS fit of the three data sets to one-species model with mass constraints (Equation (2)) (99) yields a molecular weight estimate of 25.3 ± 0.3 kDa, consistent with the predicted molecular weight of 27.4 kDa for a RecO monomer. The bottom panel shows the residuals, indicating good fits.

Inline graphic — E. coli RecO is monomeric. (A) Sedimentation velocity c(s) distribution profiles of RecO (monitored at 230 nm) at 1.5, 4 and 8 μM in buffer BTP (pH 8.0) at 25°C. (B) The weight average sedimentation coefficient of RecO slightly decreases as a function of RecO concentration, expected for a pure, non-associating species (103–106). A linear extrapolation to zero RecO concentration yields s⁰ = 0.87 ± 0.04 S. From this we calculate = 2.32 ± 0.05 S, consistent with a RecO monomer. (C) Results of sedimentation equilibrium experiments (monitored at 230 nm) performed at 4 μM RecO at three rotor speeds 20,000 (blue), 25 000 (orange) and 30 000 (gray) rpm are shown in blue, orange and gray. Each equilibrium profile is described by a single exponential. The solid lines show the best global NLLS fit of the three data sets to one-species model with mass constraints (Equation (2)) (99) yields a molecular weight estimate of 25.3 ± 0.3 kDa, consistent with the predicted molecular weight of 27.4 kDa for a RecO monomer. The bottom panel shows the residuals, indicating good fits.