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Heteronuclear solution-state NMR studies of the chromophore in cyanobacterial phytochrome Cph1.
- Source :
-
Biochemistry [Biochemistry] 2005 Jun 14; Vol. 44 (23), pp. 8244-50. - Publication Year :
- 2005
-
Abstract
- Precise structural information regarding the chromophore binding pocket is essential for an understanding of photochromicity and photoconversion in phytochrome photoreceptors. To this end, we are studying the 59 kDa N-terminal module of the cyanobacterial phytochrome Cph1 from Synechocystis sp. PCC 6803 in both thermally stable forms (Pr and Pfr) using solution-state NMR spectroscopy. The protein is deuterated, while the chromophore, phycocyanobilin (PCB), is isotopically labeled with (15)N or (13)C and (15)N. We have established a simple approach for preparing labeled PCB based on BG11 medium supplemented with an appropriate buffer and NaH(13)CO(3) and Na(15)NO(3) as sole carbon and nitrogen sources, respectively. We show that structural details of the chromophore binding pocket in both Pr and Pfr forms can be obtained using multidimensional heteronuclear solution-state NMR spectroscopy. Using one-dimensional (15)N NMR spectra, we show unequivocally that the chromophore is protonated in both Pr and Pfr states.
- Subjects :
- Bacterial Proteins metabolism
Binding Sites
Carbon Isotopes metabolism
Nitrogen Isotopes metabolism
Photochemistry methods
Photoreceptors, Microbial
Phycobilins
Phycocyanin metabolism
Phytochrome metabolism
Protein Kinases metabolism
Protons
Pyrroles metabolism
Solutions
Synechocystis metabolism
Tetrapyrroles
Thermodynamics
Bacterial Proteins chemistry
Nuclear Magnetic Resonance, Biomolecular methods
Phycocyanin chemistry
Phytochrome chemistry
Protein Kinases chemistry
Pyrroles chemistry
Synechocystis chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 0006-2960
- Volume :
- 44
- Issue :
- 23
- Database :
- MEDLINE
- Journal :
- Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 15938613
- Full Text :
- https://doi.org/10.1021/bi050457r