Skip to main content
. 2017 Sep 4;6:1636. [Version 1] doi: 10.12688/f1000research.11878.1

Figure 4. Uba1 adenylation activity as a function of Ub or ATP concentration.

Figure 4.

Kinetic assays (150µL) were run in duplicate or triplicate with 100nM WT Uba1 and XL-SMA variants in 50mM HEPES buffer at 37°C to determine initial linear rates from the first 10–20 minutes of the reaction. A) Michaelis-Menten graph of Uba1 adenylation activity as a function of Ub concentration with saturating ATP(2000nM). B) Michaelis-Menten graph of Uba1 adenylation activity as a function of ATP concentration with saturating Ub (100µM). Data was normalized to control reactions and graphed in GraphPad Prism. Km and Vmax values as well as 95% confidence intervals were also calculated in GraphPad Prism (see Table 2). Only p.E557V Km (ATP) did not have overlapping confidence intervals with WT values.