Distinct chromophore-protein environments enable asymmetric activation of a bacteriophytochrome activated diguanylate cyclase

David Buhrke, Geoffrey Gourinchas, Melanie Müller, Norbert Michael, Peter Hildebrandt, Andreas Winkler

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Sensing of red and far-red light by bacteriophytochromes involves intricate interactions between their bilin chromophore and the protein environment. The light-triggered rearrangements of the cofactor configuration and eventually the protein conformation enable bacteriophytochromes to interact with various protein effector domains for biological modulation of diverse physiological functions. Excitation of the holoproteins by red or far-red light promotes the photoconversion to their far red light-absorbing Pfr state or the red light-absorbing Pr state, respectively. Because prototypical bacteriophytochromes have a parallel dimer architecture, it is generally assumed that symmetric activation with two Pfr state protomers constitutes the signaling-active species. However, the bacteriophytochrome from Idiomarina species A28L (IsPadC) has recently been reported to enable long-range signal transduction also in asymmetric dimers containing only one Pfr protomer. By combining crystallography, hydrogen-deuterium exchange coupled to MS and vibrational spectroscopy, we show here that Pfr of IsPadC is in equilibrium with an intermediate "Pfr-like" state that combines features of Pfr and Meta-R states observed in other bacteriophytochromes. We also show that structural rearrangements in the N-terminal segment (NTS) can stabilize this Pfr-like state and that the PHY-tongue conformation of IsPadC is partially uncoupled from the initial changes in the NTS. This uncoupling enables structural asymmetry of the overall homodimeric assembly and allows signal transduction to the covalently linked physiological diguanylate cyclase output module in which asymmetry might play a role in the enzyme-catalyzed reaction. The functional differences to other phytochrome systems identified here highlight opportunities for using additional red-light sensors in artificial sensor-effector systems.

Original languageEnglish
Pages (from-to)539-551
JournalThe Journal of Biological Chemistry
Volume295
DOIs
Publication statusPublished - 10 Jan 2020

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Chromophores
Chemical activation
Light
Signal transduction
Proteins
Protein Subunits
Dimers
Conformations
Alteromonadaceae
Signal Transduction
Bile Pigments
Phytochrome
Vibrational spectroscopy
Protein Conformation
Crystallography
Deuterium
Sensors
Tongue
diguanylate cyclase
Hydrogen

Fields of Expertise

  • Human- & Biotechnology

Cite this

Distinct chromophore-protein environments enable asymmetric activation of a bacteriophytochrome activated diguanylate cyclase. / Buhrke, David; Gourinchas, Geoffrey; Müller, Melanie; Michael, Norbert; Hildebrandt, Peter; Winkler, Andreas.

In: The Journal of Biological Chemistry, Vol. 295, 10.01.2020, p. 539-551.

Research output: Contribution to journalArticleResearchpeer-review

Buhrke, David ; Gourinchas, Geoffrey ; Müller, Melanie ; Michael, Norbert ; Hildebrandt, Peter ; Winkler, Andreas. / Distinct chromophore-protein environments enable asymmetric activation of a bacteriophytochrome activated diguanylate cyclase. In: The Journal of Biological Chemistry. 2020 ; Vol. 295. pp. 539-551.
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abstract = "Sensing of red and far-red light by bacteriophytochromes involves intricate interactions between their bilin chromophore and the protein environment. The light-triggered rearrangements of the cofactor configuration and eventually the protein conformation enable bacteriophytochromes to interact with various protein effector domains for biological modulation of diverse physiological functions. Excitation of the holoproteins by red or far-red light promotes the photoconversion to their far red light-absorbing Pfr state or the red light-absorbing Pr state, respectively. Because prototypical bacteriophytochromes have a parallel dimer architecture, it is generally assumed that symmetric activation with two Pfr state protomers constitutes the signaling-active species. However, the bacteriophytochrome from Idiomarina species A28L (IsPadC) has recently been reported to enable long-range signal transduction also in asymmetric dimers containing only one Pfr protomer. By combining crystallography, hydrogen-deuterium exchange coupled to MS and vibrational spectroscopy, we show here that Pfr of IsPadC is in equilibrium with an intermediate {"}Pfr-like{"} state that combines features of Pfr and Meta-R states observed in other bacteriophytochromes. We also show that structural rearrangements in the N-terminal segment (NTS) can stabilize this Pfr-like state and that the PHY-tongue conformation of IsPadC is partially uncoupled from the initial changes in the NTS. This uncoupling enables structural asymmetry of the overall homodimeric assembly and allows signal transduction to the covalently linked physiological diguanylate cyclase output module in which asymmetry might play a role in the enzyme-catalyzed reaction. The functional differences to other phytochrome systems identified here highlight opportunities for using additional red-light sensors in artificial sensor-effector systems.",
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AU - Gourinchas, Geoffrey

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AU - Hildebrandt, Peter

AU - Winkler, Andreas

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