Real-time Monitoring of Interactions Using SPR Biosensors

The readout in a BIA experiment can be an equilibrium binding constant (JC or Kp) or, preferably, distinct association and dissociation rate constants, and respectively. This requires an experimental setup which allows monitoring of association separately from dissociation in real-time. [Pg.161]

The Model System cAMP-Dependent Protein Kinase [Pg.161]

To validate interaction assays, a suitable biological model system is helpful. cAMP-dependent protein kinase (PKA) is used as such a model system in the assays described below. PKA is the main target for the second messenger cAMP in eukaryotic cells and, since it has been implicated in various human diseases, the development of cAMP analogs modulating PKA activity has provoked increasing interest. [Pg.161]

In its inactive state, PKA is a tetrameric holoenzyme consisting of two regulatory (R, with four isoforms in humans Rp, R , Rp) and two catalytic (C, with several isoforms in humans) subunits. The binding of four molecules of cAMP to the dimeric R subunit leads to the dissociation of the holoenzyme releasing the active C subunits (Taylor et al., 2004). [Pg.161]

Real-time Monitoring of interactions Using SPR Biosensors [Pg.161]

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