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. 2023 Jul 20;51(15):8070–8084. doi: 10.1093/nar/gkad590

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© The Author(s) 2023. 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 (https://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. — N-value determination for the four LeuAC variants. (A) Single turnover time courses for the three adenine nucleotides. The green ATP + ADP curve represents the ATP consumption after accounting for ADP production and is almost a reflection of the (green) curve for AMP production. Thus, it likely represents in situ formation of leu-5′AMP. (B) Averaged n-values determined from the two physical constants, A (the amplitude of the burst) and C (the offset), with their standard deviations obtained for each variant by fitting to Eq. (1) as described in Materials and Methods. (C) Multivariate regression analysis of n-values averaged in B confirm that n-values for the appearance of ADP and AMP account almost exactly for ATP consumption. Solving the simultaneous equations [n_T_i = Σ_i,variants (β_D(C + n_D) + β_M(n_M) + ϵ)] by least squares using JMP16PRO gave the table of values underneath the graph. Here, and elsewhere, the true value of the regression line has a 95% chance of falling within the red boundaries.