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. Author manuscript; available in PMC: 2016 Feb 16.
Published in final edited form as: J Am Chem Soc. 2015 Aug 25;137(35):11303–11311. doi: 10.1021/jacs.5b04366

Figure 4.

Figure 4

P1 is fluorogenic upon binding and reacting with RA to form a covalent conjugate. P1 (5 µM) was incubated with RA (5 µM) or the K210A RA mutant (5 µM) for 24 h in buffer at 25 °C resulting in complete covalent modification of RA, except in the case of the K210A mutant which forms a noncovalent K210A RA•P1 complex. P1 (5 µM) is dark in buffer and is only very weakly fluorescent when bound to the K210A RA mutant. In contrast, P1 is strongly fluorescent upon forming a covalent conjugate with RA. Excitation and emission spectra were recorded using an Aviv fluorescence spectrometer. Samples in the inset were photographed under illumination with a hand-held UV lamp.