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. 2021 Apr 20;296:100681. doi: 10.1016/j.jbc.2021.100681

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Stopped-flow-based fluorescent BOP-Met-Leu-Leu tripeptide release kinetics by mRF1 and RF1 on RC containing cognate/near-cognate codons. The first two panels present the time courses of BOP fluorescence change due to BOP-Met-Leu-Leu tripeptide release from cognate RC_UAA and near-cognate RC_UGA (0.1 μM), with increasing concentrations of mRF1 (leftpanels: A and D) and RF1 (middlepanels: B and E) in arbitrary units (a.u.). The rightpanels (C and F) are the Michaelis–Menten plots (M-M plot) representing dependence of the observed peptide release rates (derived from the titration curves in the left and middle panels), on the concentration of mRF1 and RF1 on corresponding codons (labeled in left). The kinetic parameters of peptide release (k_cat, K_M, and k_cat/K_M) by mRF1 and RF1 are derived by fitting the curves with hyperbolic equation. The error bars indicate SEM from minimum three sets of independent experiments.