Incoherent manipulation of the photoactive yellow protein photocycle with dispersed pump-dump-probe spectroscopy

Biophys J. 2004 Sep;87(3):1858-72. doi: 10.1529/biophysj.104.043794.


Photoactive yellow protein is the protein responsible for initiating the "blue-light vision" of Halorhodospira halophila. The dynamical processes responsible for triggering the photoactive yellow protein photocycle have been disentangled with the use of a novel application of dispersed ultrafast pump-dump-probe spectroscopy, where the photocycle can be started and interrupted with appropriately tuned and timed laser pulses. This "incoherent" manipulation of the photocycle allows for the detailed spectroscopic investigation of the underlying photocycle dynamics and the construction of a fully self-consistent dynamical model. This model requires three kinetically distinct excited-state intermediates, two (ground-state) photocycle intermediates, I(0) and pR, and a ground-state intermediate through which the protein, after unsuccessful attempts at initiating the photocycle, returns to the equilibrium ground state. Also observed is a previously unknown two-photon ionization channel that generates a radical and an ejected electron into the protein environment. This second excitation pathway evolves simultaneously with the pathway containing the one-photon photocycle intermediates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Biophysical Phenomena
  • Biophysics
  • Electrons
  • Escherichia coli / metabolism
  • Halorhodospira halophila / metabolism
  • Ions
  • Kinetics
  • Light
  • Models, Statistical
  • Models, Theoretical
  • Photons
  • Photoreceptors, Microbial / chemistry*
  • Photoreceptors, Microbial / metabolism
  • Software
  • Spectrometry, Fluorescence
  • Spectrophotometry / methods*
  • Time Factors


  • Bacterial Proteins
  • Ions
  • Photoreceptors, Microbial
  • photoactive yellow protein, Bacteria