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![[Figure 1]](jb5034fig1mag.jpg)
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Figure 1
Functional properties of purified HsADA1. (a) SDS–PAGE analysis of HsADA1 before (lane 1) and after (lane 2) cleavage of the His tag with TEV protease, showing the expected decrease in molecular mass. (b) Size-exclusion chromatography elution profile for cleaved HsADA1. (c) Michaelis–Menten kinetic parameters of cleaved HsADA1 at pH 7.4, which agree with previously reported values. (d) Stability of tagged and cleaved HsADA1 obtained from differential scanning fluorimetry. The melting temperature was determined as the inflection point of the first derivative with respect to temperature of the 330 nm:350 nm absorbance ratio. (e) His-tagged HsADA1 activity as a function of pH condition (n = 32 per pH condition). Due to the different concentrations of enzyme necessary to obtain a robust signal for each pH condition, the original rate calculation (determined by linear regression from the initial 10% of substrate degradation) in units of µM adenosine per second was divided by the concentration of HsADA1 in µM to obtain a normalized unit of s−1. |
![[Figure 1]](jb5034fig1.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
