Pressure-Sensitive Reaction Yield of the TePixD Blue-Light Sensor Protein

Kuroi, Kunisato; Okajima, Koji; Ikeuchi, Masahiko; Tokutomi, Satoru; Kamiyama, Tadashi; Terazima, Masahide

doi:10.1021/jp511946u

Cited by 10 publications

(5 citation statements)

References 61 publications

(130 reference statements)

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“…The importance of the fluctuation for the dissociation reaction of the TePixD decamer was also demonstrated by highpressure TG measurement. 35 It was found that the yield of the dissociation reaction of TePixD significantly decreased by applying a relatively small pressure, although the reaction rate increased with increasing pressure. Using densimetry at high pressures, the compressibility was found to be pressure dependent even in a relatively low pressure range.…”

Section: Fluctuations Of Intermediatesmentioning

confidence: 99%

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Enhanced Conformational Fluctuations during Protein Reactions

Terazima

2019

Chem. Lett.

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Proteins are flexible systems and their structures fluctuate at room temperature in aqueous solution due to the thermal energy. Although the role of conformational fluctuations in protein reactions has been an attractive and challenging subject to elucidate reaction mechanisms, there has been no direct experimental detection of such fluctuations during reactions. In this review, several time-resolved measurements of thermodynamic properties, which are directly linked to the fluctuation of proteins, are presented. In all cases, the fluctuations determined from the thermal expansion volume and the compressibility are enhanced in short-lived intermediates, and are well correlated with the reactivities. These results show that fluctuation is indeed key to understanding why some proteins can react without significant conformational changes.

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Section: Fluctuations Of Intermediatesmentioning

confidence: 99%

“…Using densimetry at high pressures, the compressibility was found to be pressure dependent even in a relatively low pressure range. 35 It was concluded that the origin of the pressure-sensitive reaction yield is due to the decrease of compressibility at high pressure.…”

Section: Fluctuations Of Intermediatesmentioning

confidence: 99%

Enhanced Conformational Fluctuations during Protein Reactions

Terazima

2019

Chem. Lett.

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“…The experimental setup for the high-pressure TG measurement was similar to that reported before. 26,28,29 A high-pressure cell specially designed for TG measurement was used. High pressure was applied with a compact hand pump (Syn Corporation TP-501).…”

Section: Tg Measurementsmentioning

confidence: 99%

“…In the dynamic light scattering (DLS) measurements, a highpressure cell PCI-400-DLS (Syn Corporation, Japan) was used. 29 The DLS experiment was performed with a fiber-optics-based correlator (FDLS-3000, Otsuka Electronics). The sample solution was encapsulated in a high-pressure inner cell composed of quartz and silicone tubes (sample volume B300 mL), and that inner cell was placed inside the high-pressure apparatus.…”

Section: Dls Measurementsmentioning

confidence: 99%

Anomalous pressure effects on the photoreaction of a light-sensor protein from Synechocystis, PixD (Slr1694), and the compressibility change of its intermediates

Nakajima

Kuroi

Nakasone

et al. 2016

Phys. Chem. Chem. Phys.

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SyPixD (Slr1694) is a blue-light receptor that contains a BLUF (blue-light sensor using a flavin chromophore) domain for the function of phototaxis. The key reaction of this protein is a light-induced conformational change and subsequent dissociation reaction from the decamer to the dimer. In this study, anomalous effects of pressure on this reaction were discovered, and changes in the compressibility of its short-lived intermediates were investigated. While the absorption spectra of the dark and light states are not sensitive to pressure, the formation yield of the first intermediate decreases with pressure to about 40% at 150 MPa. Upon blue-light illumination with a sufficiently strong intensity, the transient grating signal, which represents the dissociation of the SyPixD decamer, was observed at 0.1 MPa, and the signal intensity significantly decreased with increasing pressure. This behavior shows that the dissociation of the decamer from the second intermediate state is suppressed by pressure. However, while the decamer undergoes no dissociation upon excitation of one monomer unit at 0.1 MPa, dissociation is gradually induced with increasing pressure. For solving this strange behavior, the compressibility changes of the intermediates were measured as a function of pressure at weak light intensity. Interestingly, the compressibility change was negative at low pressure, but became positive with increasing pressure. Because the compressibility is related to the volume fluctuation, this observation suggests that the driving force for this reaction is fluctuation of the protein. The relationship between the cavities at the interfaces of the monomer units and the reactivity was also discussed.

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“…However, since proteins are generally flexible, the protein reaction yield could be pressure dependent. 47,48 Thus, we first examined the pressure dependence of the reaction yield of the phot1LOV2-linker. The pressure effects on the UV-Vis absorption spectra in the dark state were found to be negligible after the correction of the increase in the density of the solution (i.e., concentration) ( Fig.…”

Section: Pressure Dependent Reaction Yieldmentioning

confidence: 99%

Time-resolved fluctuation during the photochemical reaction of a photoreceptor protein: phototropin1LOV2-linker

Kuroi

Sato

Nakasone

et al. 2016

Phys. Chem. Chem. Phys.

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Although the relationship between structural fluctuations and reactions is important for elucidating reaction mechanisms, experimental data describing such fluctuations of reaction intermediates are sparse. In order to investigate structural fluctuations during a protein reaction, the compressibilities of intermediate species after photoexcitation of a phot1LOV2-linker, which is a typical LOV domain protein with the C-terminal linker including the J-α helix and used recently for optogenetics, were measured in the time-domain by the transient grating and transient lens methods with a high pressure optical cell. The yield of covalent bond formation between the chromophore and a Cys residue (S state formation) relative to that at 0.1 MPa decreased very slightly with increasing pressure. The fraction of the reactive species that yields the T state (linker-unfolded state) decreased almost proportionally with pressure (0.1-200 MPa) to about 65%. Interestingly, the volume change associated with the reaction was much more pressure sensitive. By combining these data, the compressibility changes for the short lived intermediate (S state) and the final product (T state) formation were determined. The compressibility of the S state was found to increase compared with the dark (D) state, and the compressibility decreased during the transition from the S state to the T state. The compressibility change is discussed in terms of cavities inside the protein. By comparing the crystal structures of the phot1LOV2-linker at dark and light states, we concluded that the cavity volumes between the LOV domain and the linker domain increase in the S state, which explains the enhanced compressibility.

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Pressure-Sensitive Reaction Yield of the TePixD Blue-Light Sensor Protein

Cited by 10 publications

References 61 publications

Enhanced Conformational Fluctuations during Protein Reactions

Enhanced Conformational Fluctuations during Protein Reactions

Anomalous pressure effects on the photoreaction of a light-sensor protein from Synechocystis, PixD (Slr1694), and the compressibility change of its intermediates

Time-resolved fluctuation during the photochemical reaction of a photoreceptor protein: phototropin1LOV2-linker

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