| Assay Method Information | |
| | Ligand Affinity Measurements |
| Description: | Ligand binding to A. aeolicus LpxC was assayed by isothermal titration using isothermal microcalorimeter from Microcal, Inc. (Northampton, MA). The enzyme was stripped of all metal ions, and was reconstituted with Zn2+ (1:1 ratio) by the addition of ZnSO4. The calorimeter cell contained enzyme (40 or 60 uM), and the syringe contained ligand (250 or 400 uM). A series of 30 injections of 8 ul each were performed at 180-s intervals. Titrations of aliphatic compounds into buffer were also performed as control experiments using identical conditions. Data were fit to a single binding site model using Origin v. 2.9. |
| Affinity data for this assay | |
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