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Title: Photoreversible interconversion of a phytochrome photosensory module in the crystalline state

Abstract

A major barrier to defining the structural intermediates that arise during the reversible photointerconversion of phytochromes between their biologically inactive and active states has been the lack of crystals that faithfully undergo this transition within the crystal lattice. Here, we describe a crystalline form of the cyclic GMP phosphodiesterases/adenylyl cyclase/FhlA (GAF) domain from the cyanobacteriochrome PixJ in Thermosynechococcus elongatus assembled with phycocyanobilin that permits reversible photoconversion between the blue light-absorbing Pb and green light-absorbing Pg states, as well as thermal reversion of Pg back to Pb. The X-ray crystallographic structure of Pb matches previous models, including autocatalytic conversion of phycocyanobilin to phycoviolobilin upon binding and its tandem thioether linkage to the GAF domain. Cryocrystallography at 150 K, which compared diffraction data from a single crystal as Pb or after irradiation with blue light, detected photoconversion product(s) based on F obs- F obs difference maps that were consistent with rotation of the bonds connecting pyrrole rings C and D. Further spectroscopic analyses showed that phycoviolobilin is susceptible to X-ray radiation damage, especially as Pg, during single-crystal X-ray diffraction analyses, which could complicate fine mapping of the various intermediate states. Fortunately, we found that PixJ crystals are amenable to serial femtosecond crystallographymore » (SFX) analyses using X-ray free-electron lasers (XFELs). As proof of principle, we solved by room temperature SFX the GAF domain structure of Pb to 1.55-Å resolution, which was strongly congruent with synchrotron-based models. Analysis of these crystals by SFX should now enable structural characterization of the early events that drive phytochrome photoconversion.« less

Authors:
ORCiD logo [1];  [2];  [2];  [3];  [4];  [4];  [5];  [3];  [3];  [3];  [3];  [3];  [3];  [3];  [3]; ORCiD logo [5];  [5];  [5];  [5];  [3] more »;  [3];  [3];  [6]; ORCiD logo [3]; ORCiD logo [7];  [2]; ORCiD logo [1] « less
  1. Washington Univ., St. Louis, MO (United States)
  2. Rice Univ., Houston, TX (United States)
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  4. Diamond Light Source Ltd., Oxfordshire (United Kingdom)
  5. SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source
  6. SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource
  7. Diamond Light Source Ltd., Oxfordshire (United Kingdom); Rutherford Appleton Lab., Didcot, Oxfordshire (United Kingdom)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Institutes of Health (NIH); National Science Foundation (NSF); Diamond Light Source; Biotechnology and Biological Sciences Research Council; Wellcome Trust
OSTI Identifier:
1599832
Alternate Identifier(s):
OSTI ID: 1604574
Grant/Contract Number:  
AC02-05CH11231; GM127892; GM055302; GM110501; GM126289; GM117126; 1231306; 102593; AC02-76SF00515; AC02-06CH11357; 085P1000817; ACB-12002; AGM-12006; 1S10OD012289-01A1
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 117; Journal Issue: 1; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; phytochrome; photoreceptor; X-ray crystallography

Citation Formats

Burgie, E. Sethe, Clinger, Jonathan A., Miller, Mitchell D., Brewster, Aaron S., Aller, Pierre, Butryn, Agata, Fuller, Franklin D., Gul, Sheraz, Young, Iris D., Pham, Cindy C., Kim, In-Sik, Bhowmick, Asmit, O’Riordan, Lee J., Sutherlin, Kyle D., Heinemann, Joshua V., Batyuk, Alexander, Alonso-Mori, Roberto, Hunter, Mark S., Koglin, Jason E., Yano, Junko, Yachandra, Vittal K., Sauter, Nicholas K., Cohen, Aina E., Kern, Jan, Orville, Allen M., Phillips, George N., and Vierstra, Richard D. Photoreversible interconversion of a phytochrome photosensory module in the crystalline state. United States: N. p., 2019. Web. doi:10.1073/pnas.1912041116.
Burgie, E. Sethe, Clinger, Jonathan A., Miller, Mitchell D., Brewster, Aaron S., Aller, Pierre, Butryn, Agata, Fuller, Franklin D., Gul, Sheraz, Young, Iris D., Pham, Cindy C., Kim, In-Sik, Bhowmick, Asmit, O’Riordan, Lee J., Sutherlin, Kyle D., Heinemann, Joshua V., Batyuk, Alexander, Alonso-Mori, Roberto, Hunter, Mark S., Koglin, Jason E., Yano, Junko, Yachandra, Vittal K., Sauter, Nicholas K., Cohen, Aina E., Kern, Jan, Orville, Allen M., Phillips, George N., & Vierstra, Richard D. Photoreversible interconversion of a phytochrome photosensory module in the crystalline state. United States. doi:10.1073/pnas.1912041116.
Burgie, E. Sethe, Clinger, Jonathan A., Miller, Mitchell D., Brewster, Aaron S., Aller, Pierre, Butryn, Agata, Fuller, Franklin D., Gul, Sheraz, Young, Iris D., Pham, Cindy C., Kim, In-Sik, Bhowmick, Asmit, O’Riordan, Lee J., Sutherlin, Kyle D., Heinemann, Joshua V., Batyuk, Alexander, Alonso-Mori, Roberto, Hunter, Mark S., Koglin, Jason E., Yano, Junko, Yachandra, Vittal K., Sauter, Nicholas K., Cohen, Aina E., Kern, Jan, Orville, Allen M., Phillips, George N., and Vierstra, Richard D. Wed . "Photoreversible interconversion of a phytochrome photosensory module in the crystalline state". United States. doi:10.1073/pnas.1912041116. https://www.osti.gov/servlets/purl/1599832.
@article{osti_1599832,
title = {Photoreversible interconversion of a phytochrome photosensory module in the crystalline state},
author = {Burgie, E. Sethe and Clinger, Jonathan A. and Miller, Mitchell D. and Brewster, Aaron S. and Aller, Pierre and Butryn, Agata and Fuller, Franklin D. and Gul, Sheraz and Young, Iris D. and Pham, Cindy C. and Kim, In-Sik and Bhowmick, Asmit and O’Riordan, Lee J. and Sutherlin, Kyle D. and Heinemann, Joshua V. and Batyuk, Alexander and Alonso-Mori, Roberto and Hunter, Mark S. and Koglin, Jason E. and Yano, Junko and Yachandra, Vittal K. and Sauter, Nicholas K. and Cohen, Aina E. and Kern, Jan and Orville, Allen M. and Phillips, George N. and Vierstra, Richard D.},
abstractNote = {A major barrier to defining the structural intermediates that arise during the reversible photointerconversion of phytochromes between their biologically inactive and active states has been the lack of crystals that faithfully undergo this transition within the crystal lattice. Here, we describe a crystalline form of the cyclic GMP phosphodiesterases/adenylyl cyclase/FhlA (GAF) domain from the cyanobacteriochrome PixJ in Thermosynechococcus elongatus assembled with phycocyanobilin that permits reversible photoconversion between the blue light-absorbing Pb and green light-absorbing Pg states, as well as thermal reversion of Pg back to Pb. The X-ray crystallographic structure of Pb matches previous models, including autocatalytic conversion of phycocyanobilin to phycoviolobilin upon binding and its tandem thioether linkage to the GAF domain. Cryocrystallography at 150 K, which compared diffraction data from a single crystal as Pb or after irradiation with blue light, detected photoconversion product(s) based on Fobs- Fobs difference maps that were consistent with rotation of the bonds connecting pyrrole rings C and D. Further spectroscopic analyses showed that phycoviolobilin is susceptible to X-ray radiation damage, especially as Pg, during single-crystal X-ray diffraction analyses, which could complicate fine mapping of the various intermediate states. Fortunately, we found that PixJ crystals are amenable to serial femtosecond crystallography (SFX) analyses using X-ray free-electron lasers (XFELs). As proof of principle, we solved by room temperature SFX the GAF domain structure of Pb to 1.55-Å resolution, which was strongly congruent with synchrotron-based models. Analysis of these crystals by SFX should now enable structural characterization of the early events that drive phytochrome photoconversion.},
doi = {10.1073/pnas.1912041116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 1,
volume = 117,
place = {United States},
year = {2019},
month = {12}
}

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