We obtained the absorption spectrum of the N intermediate that plays an important role in the proton uptake during the photocycle of bacteriorhodopsin. The reported spectra of N that were reconstituted from the transient difference absorption spectrum depended on the assumed kinetic models such as sequential M f N f O reaction and M a N a O quasi-equilibrium. In this work we measured the excitation spectrum of N by monitoring the characteristic absorption of Q (a photoproduct of N) at 664 nm. The λ max and max of N were 559 ( 3 nm and (4.30 ( 0.75) × 10 4 M -1 cm -1 , respectively. The present spectrum is consistent with the results of the recent SVD analysis. This work provides independent evidence that Q is photochemically formed from N.
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