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Optical control of protein-protein interactions via blue light-induced domain swapping.

ABSTRACT: The design of new optogenetic tools for controlling protein function would be facilitated by the development of protein scaffolds that undergo large, well-defined structural changes upon exposure to light. Domain swapping, a process in which a structural element of a monomeric protein is replaced by the same element of another copy of the same protein, leads to a well-defined change in protein structure. We observe domain swapping in a variant of the blue light photoreceptor photoactive yellow protein in which a surface loop is replaced by a well-characterized protein-protein interaction motif, the E-helix. In the domain-swapped dimer, the E-helix sequence specifically binds a partner K-helix sequence, whereas in the monomeric form of the protein, the E-helix sequence is unable to fold into a binding-competent conformation and no interaction with the K-helix is seen. Blue light irradiation decreases the extent of domain swapping (from Kd = 10 ?M to Kd = 300 ?M) and dramatically enhances the rate, from weeks to <1 min. Blue light-induced domain swapping thus provides a novel mechanism for controlling of protein-protein interactions in which light alters both the stability and the kinetic accessibility of binding-competent states.

SUBMITTER: Reis JM

PROVIDER: S-EPMC4372075 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature

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Optical control of protein-protein interactions via blue light-induced domain swapping.

Reis Jakeb M JM Burns Darcy C DC Woolley G Andrew GA

Biochemistry 20140716 30

The design of new optogenetic tools for controlling protein function would be facilitated by the development of protein scaffolds that undergo large, well-defined structural changes upon exposure to light. Domain swapping, a process in which a structural element of a monomeric protein is replaced by the same element of another copy of the same protein, leads to a well-defined change in protein structure. We observe domain swapping in a variant of the blue light photoreceptor photoactive yellow p ...[more]

PMID: 25003701

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Project description:PURPOSES:To evaluate the optical performance of blue-light filtering spectacle lenses and investigate whether a reduction in blue light transmission affects visual performance and sleep quality. METHODS:Experiment 1: The relative changes in phototoxicity, scotopic sensitivity, and melatonin suppression of five blue-light filtering plano spectacle lenses were calculated based on their spectral transmittances measured by a spectrophotometer. Experiment 2: A pseudo-randomized controlled study was conducted to evaluate the clinical performance of two blue-light filtering spectacle lenses (BF: blue-filtering anti-reflection coating; BT: brown-tinted) with a regular clear lens (AR) serving as a control. A total of eighty computer users were recruited from two age cohorts (young adults: 18-30 yrs, middle-aged adults: 40-55 yrs). Contrast sensitivity under standard and glare conditions, and colour discrimination were measured using standard clinical tests. After one month of lens wear, subjective ratings of lens performance were collected by questionnaire. RESULTS:All tested blue-light filtering spectacle lenses theoretically reduced the calculated phototoxicity by 10.6% to 23.6%. Although use of the blue-light filters also decreased scotopic sensitivity by 2.4% to 9.6%, and melatonin suppression by 5.8% to 15.0%, over 70% of the participants could not detect these optical changes. Our clinical tests revealed no significant decrease in contrast sensitivity either with (95% confidence intervals [CI]: AR-BT [-0.05, 0.05]; AR-BF [-0.05, 0.06]; BT-BF [-0.06, 0.06]) or without glare (95% CI: AR-BT [-0.01, 0.03]; AR-BF [-0.01, 0.03]; BT-BF [-0.02, 0.02]) and colour discrimination (95% CI: AR-BT [-9.07, 1.02]; AR-BF [-7.06, 4.46]; BT-BF [-3.12, 8.57]). CONCLUSION:Blue-light filtering spectacle lenses can partially filter high-energy short-wavelength light without substantially degrading visual performance and sleep quality. These lenses may serve as a supplementary option for protecting the retina from potential blue-light hazard. TRIAL REGISTRATION:ClinicalTrials.gov NCT02821403.

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Optical control of protein-protein interactions via blue light-induced domain swapping.

Publications

Optical control of protein-protein interactions via blue light-induced domain swapping.

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