A light-sensing knot revealed by the structure of the chromophore-binding domain of phytochrome

Phytochromes are red/far-red light photoreceptors that direct photosensory responses across the bacterial, fungal and plant kingdoms. These include photosynthetic potential and pigmentation in bacteria as well as chloroplast development and photomorphogenesis in plants. Phytochromes consist of an amino-terminal region that covalently binds a single bilin chromophore, followed by a carboxy-terminal dimerization domain that often transmits the light signal through a histidine kinase relay. Here we describe the three-dimensional structure of the chromophore-binding domain of Deinococcus radiodurans phytochrome assembled with its chromophore biliverdin in the Pr ground state. Our model, refined to 2.5 Å resolution, reaffirms Cys 24 as the chromophore attachment site, locates key amino acids that form a solvent-shielded bilin-binding pocket, and reveals an unusually formed deep trefoil knot that stabilizes this region. The structure provides the first three-dimensional glimpse into the photochromic behaviour of these photoreceptors and helps to explain the evolution of higher plant phytochromes from prokaryotic precursors.

[1]  R. Vierstra,et al.  Bacteriophytochromes: phytochrome-like photoreceptors from nonphotosynthetic eubacteria. , 1999, Science.

[2]  J. Lagarias,et al.  Chromopeptides from phytochrome. The structure and linkage of the PR form of the phytochrome chromophore , 1980 .

[3]  C. Litterst,et al.  Structure of the NCoA-1/SRC-1 PAS-B domain bound to the LXXLL motif of the STAT6 transactivation domain. , 2004, Journal of molecular biology.

[4]  Robert D. Finn,et al.  The Pfam protein families database , 2004, Nucleic Acids Res..

[5]  S. Golden,et al.  Biochemical Properties of CikA, an Unusual Phytochrome-like Histidine Protein Kinase That Resets the Circadian Clock in Synechococcus elongatus PCC 7942* , 2003, Journal of Biological Chemistry.

[6]  P. Hildebrandt,et al.  Protonation state and structural changes of the tetrapyrrole chromophore during the Pr --> Pfr phototransformation of phytochrome: a resonance Raman spectroscopic study. , 1999, Biochemistry.

[7]  David M. Blow,et al.  Microbatch crystallization under oil — a new technique allowing many small-volume crystallization trials , 1992 .

[8]  Thomas C Terwilliger,et al.  Automated structure solution, density modification and model building. , 2002, Acta crystallographica. Section D, Biological crystallography.

[9]  William R. Taylor,et al.  Protein knots: A tangled problem , 2003, Nature.

[10]  Winslow R. Briggs,et al.  Handbook of Photosensory Receptors , 2005 .

[11]  William R. Taylor,et al.  A deeply knotted protein structure and how it might fold , 2000, Nature.

[12]  John N Moore,et al.  A Resonance Raman Spectroscopic Study of Bilirubin - Serum Albumin Complexes , 1993 .

[13]  L. Johnson Macromolecular crystallography. Part A. Methods in Enzymology, Vol. 276. Edited by Charles W. Carter Jr and Robert M. Sweet. New York: Academic Press, 1997. Pp. xxxii + 700. Price $99.00. ISBN 0-12-182177-3 , 1998 .

[14]  Seth J. Davis,et al.  Bacteriophytochromes are photochromic histidine kinases using a biliverdin chromophore , 2001, Nature.

[15]  K. Yeh,et al.  A cyanobacterial phytochrome two-component light sensory system. , 1997, Science.

[16]  T. Lamparter,et al.  The biliverdin chromophore binds covalently to a conserved cysteine residue in the N-terminus of Agrobacterium phytochrome Agp1. , 2004, Biochemistry.

[17]  K. Uchida,et al.  In vitro assembly of phytochrome B apoprotein with synthetic analogs of the phytochrome chromophore , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Grossman,et al.  RcaE is a complementary chromatic adaptation photoreceptor required for green and red light responsiveness , 2004, Molecular microbiology.

[19]  M. Ikeuchi,et al.  Cyanobacterial phytochrome-like PixJ1 holoprotein shows novel reversible photoconversion between blue- and green-absorbing forms. , 2004, Plant & cell physiology.

[20]  S J Wodak,et al.  SFCHECK: a unified set of procedures for evaluating the quality of macromolecular structure-factor data and their agreement with the atomic model. , 1999, Acta crystallographica. Section D, Biological crystallography.

[21]  Peter Hildebrandt,et al.  Determination of the chromophore structures in the photoinduced reaction cycle of phytochrome. , 2004, Journal of the American Chemical Society.

[22]  P. Sassone-Corsi,et al.  Crystal structure and interactions of the PAS repeat region of the Drosophila clock protein PERIOD. , 2005, Molecular cell.

[23]  J. Lagarias,et al.  Harnessing phytochrome's glowing potential. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Shigeyuki Yokoyama,et al.  An enzyme with a deep trefoil knot for the active-site architecture. , 2002, Acta crystallographica. Section D, Biological crystallography.

[25]  Anastassis Perrakis,et al.  Automated protein model building combined with iterative structure refinement , 1999, Nature Structural Biology.

[26]  P. Quail,et al.  Phytochrome photosensory signalling networks , 2002, Nature Reviews Molecular Cell Biology.

[27]  S. P. Fodor,et al.  Resonance Raman analysis of the Pr and Pfr forms of phytochrome. , 1990, Biochemistry.

[28]  R. Vierstra,et al.  Phytochromes in Microorganisms , 2005 .

[29]  R. Vierstra,et al.  Carboxy-terminal deletion analysis of oat phytochrome A reveals the presence of separate domains required for structure and biological activity. , 1993, The Plant cell.

[30]  G Bricogne,et al.  Generation, representation and flow of phase information in structure determination: recent developments in and around SHARP 2.0. , 2003, Acta crystallographica. Section D, Biological crystallography.

[31]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[32]  R. Vierstra,et al.  Phylogenetic analysis of the phytochrome superfamily reveals distinct microbial subfamilies of photoreceptors. , 2005, The Biochemical journal.

[33]  R. Huber,et al.  Isolation, crystallization, crystal structure analysis and refinement of constitutive C-phycocyanin from the chromatically adapting cyanobacterium Fremyella diplosiphon at 1.66 A resolution. , 1991, Journal of molecular biology.

[34]  D E McRee,et al.  XtalView/Xfit--A versatile program for manipulating atomic coordinates and electron density. , 1999, Journal of structural biology.

[35]  Youngchang Kim,et al.  Deep trefoil knot implicated in RNA binding found in an archaebacterial protein , 2002, Proteins.

[36]  J. Chory,et al.  Regulation of Phytochrome B Nuclear Localization through Light-Dependent Unmasking of Nuclear-Localization Signals , 2005, Current Biology.

[37]  S. Steinbacher,et al.  Isolation, crystallization, crystal structure analysis and refinement of allophycocyanin from the cyanobacterium Spirulina platensis at 2.3 A resolution. , 1995, Journal of molecular biology.

[38]  Sophie E Jackson,et al.  Folding studies on a knotted protein. , 2005, Journal of molecular biology.

[39]  Koji Shirai,et al.  Biliverdin Binds Covalently to Agrobacterium Phytochrome Agp1 via Its Ring A Vinyl Side Chain* , 2003, Journal of Biological Chemistry.

[40]  T. Hirano,et al.  PHYTOCHROME PHOTOCHROMISM PROBED BY SITE-DIRECTED MUTATIONS AND CHROMOPHORE ESTERIFICATION , 1997 .

[41]  G N Murshudov,et al.  Use of TLS parameters to model anisotropic displacements in macromolecular refinement. , 2001, Acta crystallographica. Section D, Biological crystallography.

[42]  T. Lamparter,et al.  Sterically Locked Synthetic Bilin Derivatives and Phytochrome Agp1 from Agrobacterium tumefaciens Form Photoinsensitive Pr- and Pfr-like Adducts* , 2005, Journal of Biological Chemistry.

[43]  P. Adams,et al.  Substructure search procedures for macromolecular structures. , 2003, Acta crystallographica. Section D, Biological crystallography.

[44]  V. Ramakrishnan,et al.  Crystal structure of globular domain of histone H5 and its implications for nucleosome binding , 1993, Nature.

[45]  Harry Smith Physiological and Ecological Function within the Phytochrome Family , 1995 .

[46]  Shu-Hsing Wu,et al.  Defining the bilin lyase domain: lessons from the extended phytochrome superfamily. , 2000, Biochemistry.

[47]  J. Ecker,et al.  Phytochrome-Specific Type 5 Phosphatase Controls Light Signal Flux by Enhancing Phytochrome Stability and Affinity for a Signal Transducer , 2005, Cell.

[48]  Thomas Terwilliger,et al.  SOLVE and RESOLVE: automated structure solution, density modification and model building. , 2004, Journal of synchrotron radiation.

[49]  David Eisenberg,et al.  3D domain swapping: As domains continue to swap , 2002, Protein science : a publication of the Protein Society.