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Assay Method Information

Assay Name:  Peptidyl Prolyl Isomerase (PPIase) Inibition Assay
Description:  PPIase(Rotamase) activity of FKBP12 was assayed using the peptide N-succinyl Ala-Leu-Pro-Phe p-nitroanilide as substrate. It is based on the observation that chymotrypsin cleaves the C-terminal amide bond only in the trans conformer of the chromogenic substrate. The rapid hydrolysis perturbs the cis-trans conformational equilibrium, which allows one to monitor the PPIase-catalyzed cis-to-trans isomerization. Absorbance readings were collected on a spectrophotometer by use of commercial data acquisition software. The progress curves were analyzed by nonlinear least-squares fit to the integrated rate equation.
Affinity data for this assay
 

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Last update November 1, 2007
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