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![[Figure 3]](qh5068fig3mag.jpg)
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Figure 3
Validation of the substrate binding in ASBTYf by microscale thermophoresis (MST). (a) Example of a single Kd measurement of ASBTYf-Pair3 for the substrate TCA by MST. The # symbol indicates the ASBTYf-Pair3 sample before disulfide cross-linking. (b) Kd determination of ASBTYf variants for TCA by MST. Compared with wild-type (WT) ASBTYf, ASBTYf-Pair3# has a p-value of >0.05 and the T130A, N259A and H286A mutants have p-values of <0.05, while all other mutants have p-values of <0.01 (Student's t-test). (c) Docking models for TCA (cyan), GCA (blue) and CHA (pink) in the outward-facing PDB entry 4n7x structure. The distances between the sulfonic group of the TCA tail and His286 or Asn287 are indicated by blue or red numbers, respectively. (d) Kd measurements of WT ASBTYf and H286A and N287V mutants for TCA, GCA and CHA. For each substrate, the mutant groups were compared with the wild-type group and Student's t-test was performed. *, p < 0.05. For (b) and (d), all measurements were performed in four independently repeated experiments (N = 4) and the results were expressed as mean ± SD. |
![[Figure 3]](qh5068fig3.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
