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Ultrafast dynamics of phytochrome from the cyanobacterium synechocystis, reconstituted with phycocyanobilin and phycoerythrobilin.
- Source :
-
Biophysical journal [Biophys J] 2002 Feb; Vol. 82 (2), pp. 1004-16. - Publication Year :
- 2002
-
Abstract
- Femtosecond time-resolved transient absorption spectroscopy was employed to characterize for the first time the primary photoisomerization dynamics of a bacterial phytochrome system in the two thermally stable states of the photocycle. The 85-kDa phytochrome Cph1 from the cyanobacterium Synechocystis PCC 6803 expressed in Escherichia coli was reconstituted with phycocyanobilin (Cph1-PCB) and phycoerythrobilin (Cph1-PEB). The red-light-absorbing form Pr of Cph1-PCB shows an approximately 150 fs relaxation in the S(1) state after photoexcitation at 650 nm. The subsequent Z-E isomerization between rings C and D of the linear tetrapyrrole-chromophore is best described by a distribution of rate constants with the first moment at (16 ps)(-1). Excitation at 615 nm leads to a slightly broadened distribution. The reverse E-Z isomerization, starting from the far-red-absorbing form Pfr, is characterized by two shorter time constants of 0.54 and 3.2 ps. In the case of Cph1-PEB, double-bond isomerization does not take place, and the excited-state lifetime extends into the nanosecond regime. Besides a stimulated emission rise time between 40 and 150 fs, no fast relaxation processes are observed. This suggests that the chromophore-protein interaction along rings A, B, and C does not contribute much to the picosecond dynamics observed in Cph1-PCB but rather the region around ring D near the isomerizing C(15) [double bond] C(16) double bond. The primary reaction dynamics of Cph1-PCB at ambient temperature is found to exhibit very similar features as those described for plant type A phytochrome, i.e., a relatively slow Pr, and a fast Pfr, photoreaction. This suggests that the initial reactions were established already before evolution of plant phytochromes began.
- Subjects :
- Biophysical Phenomena
Biophysics
Carbon chemistry
Cysteine chemistry
Escherichia coli metabolism
Kinetics
Light
Models, Chemical
Models, Statistical
Phycobilins
Plants metabolism
Protein Conformation
Spectrophotometry
Tetrapyrroles
Time Factors
Cyanobacteria chemistry
Phycocyanin chemistry
Phycoerythrin chemistry
Phytochrome chemistry
Pyrroles chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 0006-3495
- Volume :
- 82
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Biophysical journal
- Publication Type :
- Academic Journal
- Accession number :
- 11806940
- Full Text :
- https://doi.org/10.1016/S0006-3495(02)75460-X