Photoreaction of BlrP1: the role of a nonlinear photo-intensity sensor

Shibata, Kayoko; Nakasone, Yusuke; Terazima, Masahide

doi:10.1039/c7cp08436f

Cited by 18 publications

(38 citation statements)

References 34 publications

(53 reference statements)

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“…The procedure for expression and purification of the blrP1 gene has been described previously [7]. Briefly, the blrP1 gene (N559_2724) was cloned into the pET expression vector pET His6 MBP TEV LIC (2 M‐T), which was a gift from Scott Gradia (Addgene plasmid #29708).…”

Section: Methodsmentioning

confidence: 99%

“…It has been suggested that this change allosterically regulates the enzymatic activity through the interaction at the dimeric interface [5,6]. Recently, the reaction scheme and kinetics of BlrP1 have been identified by the transient grating (TG) method [7]. The conformational change was detected as a significant decrease in its diffusion coefficient (D), which was mainly attributed to the quaternary structural change in the dimer via a diffusion-sensitive conformational change (DSCC).…”

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confidence: 99%

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Enzymatic activity of the blue light‐regulated phosphodiesterase BlrP1 from Klebsiella pneumoniae shows a nonlinear dependence on light intensity

2021

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The blue light‐regulated phosphodiesterase BlrP1 from Klebsiella pneumoniae hydrolyzes cyclic dimeric guanosine monophosphate (GMP) in a blue light‐dependent manner. It contains a photosensing BLUF domain and a functional EAL domain. Previously, it was reported that conformational changes in the dimer upon light illumination occurred only when both protomers of the dimer were excited. Based on this observation, it was proposed that BlrP1 might be a nonlinear light intensity sensor. To test this, here, the correlation between the turnover number of the hydrolysis reaction (kcat) and the fraction of the excited protein (fred) was measured by simultaneously monitoring the reaction rate and fred. Our results show that kcat is proportional to fred2. Thus, BlrP1 works as a nonlinear light intensity sensor to sense a strong light environment.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Enzymatic activity of the blue light‐regulated phosphodiesterase BlrP1 from Klebsiella pneumoniae shows a nonlinear dependence on light intensity

2021

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“…Photon absorption induces structural changes in the BLUF domain that are propagated to an adjacent output domain to regulate it allosterically. A detailed mechanism of sensory perception by the BLUF domain-containing c-di-GMP-specific PDE BlrP1 ( 26 , 46 , 93 – 97 ) is discussed in the section on photosensing.…”

Section: Modular Sensory and Receiver Domains Found In Dgcs And C-di-...mentioning

confidence: 99%

Sensory Perception in Bacterial Cyclic Diguanylate Signal Transduction

et al. 2022

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Cyclic diguanylate (c-di-GMP) signal transduction systems provide bacteria the ability to sense changing cell status or environmental conditions and then execute suitable physiological and social behaviours in response. In this review, we provide a comprehensive census of the stimuli and receptors that are linked to modulation of intracellular c-di-GMP. Emerging evidence indicates that c-di-GMP networks sense light, surfaces, energy, redox potential, respiratory electron acceptors, temperature, and structurally diverse biotic and abiotic chemicals. Bioinformatic analysis of sensory domains in diguanylate cyclases and c-di-GMP-specific phosphodiesterases as well as the receptor complexes associated with them reveals that these functions are linked to a diverse repertoire of protein domain families. We describe the principles of stimulus perception learned from studying these modular sensory devices, illustrate how they are assembled in varied combinations with output domains, and summarize a system for classifying these sensor proteins based on their complexity. Biological information-processing via c-di-GMP signal transduction is not only fundamental to bacterial survival in dynamic environments, but also is being used to engineer gene expression circuitry and synthetic proteins with à la carte biochemical functionalities.

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“…Previous transient grating (TG) measurements have revealed that the two proteins (BlrP1 and YcgF) also show millisecond conformational changes, despite the absence of Trp and the τ2-phase. 37,38 Hence, in these cases, the slow conformational change may be caused by the τ1-phase through different signaling pathways. NMR measurements of BlrP1 indicate that the blue light induces conformational changes in α3-and α4-helices extending from the Cterminus of the BLUF domain.…”

Section: Origin Of τ 2 -Phasementioning

confidence: 99%

“…Previously, the global conformational changes of many BLUF proteins have been detected by the TG technique. 37,38,43,44,45 For example, the time constants of the conformation changes far from the chromophore for PapB, BlrP1, SyPixD, and TePixD have been reported to be 24 ms, 45 21 ms, 38 350 ms, 46 and 4 ms, 44 , respectively. It is interesting to note that the time constant of PapB (24 ms) are relatively close to those observed from the slow absorption change (τ2 = 12 ms).…”

Section: Signaling Pathway Of Bluf Proteinsmentioning

confidence: 99%

Slow conformational changes of blue light sensor BLUF proteins in milliseconds

Tokonami

Onose

Nakasone

et al. 2021

Preprint

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BLUF (blue light sensor using flavin) proteins consist of flavin-binding BLUF domains and functional domains. Upon blue light excitation, the hydrogen bond network around the flavin chromophore changes, and the absorption spectrum in the visible region exhibits red-shift. Ultimately, the light information received in the BLUF domain is transmitted to the functional region. It has been believed that this red-shift is complete within nanoseconds. Contrary to this commonly accepted scheme, in this study, slow reaction kinetics were discovered in milliseconds (τ1- and τ2-phase) for all the BLUF proteins examined (AppA, OaPAC, BlrP1, YcgF, PapB, SyPixD, and TePixD). Despite extensive reports on BLUF, this is the first clear observation of the BLUF protein absorption change with the duration in the millisecond time region. From the measurements of some domain-deleted mutants of OaPAC and two chimeric mutants of PixD proteins, it was found that the slower dynamics (τ2-phase) are strongly affected by the size and nature of the C-terminal region adjacent to the BLUF domain. Hence, this millisecond reaction is a significant indicator of conformational changes in the C-terminal region, which is essential for the biological functions. On the other hand, the τ1-phase commonly exists in all BLUF proteins, including any mutants. The origin of the slow dynamics was studied using site-specific mutants. These results clearly show the importance of Trp in the BLUF domain. Based on this, a reaction scheme for the BLUF reaction is proposed.

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Photoreaction of BlrP1: the role of a nonlinear photo-intensity sensor

Cited by 18 publications

References 34 publications

Enzymatic activity of the blue light‐regulated phosphodiesterase BlrP1 from Klebsiella pneumoniae shows a nonlinear dependence on light intensity

Enzymatic activity of the blue light‐regulated phosphodiesterase BlrP1 from Klebsiella pneumoniae shows a nonlinear dependence on light intensity

Sensory Perception in Bacterial Cyclic Diguanylate Signal Transduction

Slow conformational changes of blue light sensor BLUF proteins in milliseconds

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