Design and engineering of light-sensitive protein switches

McCue, Amelia C.; Kuhlman, Brian

doi:10.1016/j.sbi.2022.102377

Cited by 9 publications

(6 citation statements)

References 66 publications

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“…Other novel optogenetic systems have also been developed by researchers for optogenetic gene expression. The photosensitive molecules in optogenetic systems often contain several components of large size and multidomain structure [191] . In 2021, Verkhusha et al.…”

Section: The Application Of Nir Light For Neuromodulationmentioning

confidence: 99%

“…The photosensitive molecules in optogenetic systems often contain several components of large size and multidomain structure. [191] In 2021, Verkhusha et al developed a single-component NIR-inducible optogenetic system by engineering photosensitive constructs optimized IsPadC-PCM (photosensory core module of Idiomarina sp. A28L phytochrome-activated diguanylyl cyclase) variant, named iLight, which is the minimal light-sensing module in photochromes (Figure 12b).…”

Section: The Gene Expression Of Neurons Mediated By Nirmentioning

confidence: 99%

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Advanced near‐infrared light approaches for neuroimaging and neuromodulation

Yin

Jiang

Liu

et al. 2023

BMEMat

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Almost all physiological processes of animals are controlled by the brain, including language, cognitive, memory, learning, emotion and so forth. Minor brain dysfunction usually leads to brain diseases and disorders. Therefore, it' is greatly meaningful and urgent for scientists to have a better understanding of brain structure and function. Optical approaches can provide powerful tools for imaging and modulating physiological processes of the brain. In particular, optical approaches in the near-infrared (NIR) window (700-1700 nm) exhibit excellent prosperities of deep tissue penetration and low tissue scattering and absorption compared with those of visible windows (400-700 nm), which provides a promising approach for scientists to develop desired methods of neuroimaging and neuromodulation in deep brain tissues. In this review, variable types of NIR light approaches for imaging and modulating neural ions, membrane potential, neurotransmitters, and other critical molecules for brain functions and diseases are summarized. In particular, the latest breakthrough research of brain imaging and brain regulation in the NIR-II window (1000-1700 nm) are highlighted. Finally, we conclude the challenges and prospects of NIR light-based neuroimaging and neuromodulation for both basic brain research and further clinical translation.

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Section: The Application Of Nir Light For Neuromodulationmentioning

confidence: 99%

Section: The Gene Expression Of Neurons Mediated By Nirmentioning

confidence: 99%

Advanced near‐infrared light approaches for neuroimaging and neuromodulation

Yin

Jiang

Liu

et al. 2023

BMEMat

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“…LOV domain proteins provide a novel platform owing to their small molecular size and quick response time. 3 These tools take advantage of the fluorescent properties of LOV proteins to develop fluorescence imaging tools and possible implementation in super-resolution microscopy. 4 The LOV-based fluorescent reporter systems have shown increased brightness, improved stability and functionality in low-oxygen and anaerobic conditions, compared to the green fluorescent proteins.…”

Section: Introductionmentioning

confidence: 99%

“…The photochemistry of the LOV domain is triggered when the chromophore absorbs light and attains a singlet excited state (S 1 ), followed by intersystem crossing to a triplet state (T 1 ). The process induces a proton transfer to the N5 atom, changing the hybridization of the C4a atom to sp 3 , and generates a tilt in the planarity of the isoalloxazine ring. This photocycle mediated by a radical-pair mechanism involve the thiol group of a conserved cysteine and the C4a atom of flavin, enabling electron transfer and subsequent formation of a covalent adduct.…”

Section: Introductionmentioning

confidence: 99%

Network analysis of chromophore binding site in LOV domain

Panda

Krishnamoorthy

et al. 2022

Preprint

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Photoreceptor proteins are versatile toolboxes for developing biosensors for optogenetic applications. These molecular tools get activated upon illumination of blue light, which in turn offers a non-invasive method for gaining high spatiotemporal resolution and precise control of cellular signal transduction. The Light-Oxygen-Voltage (LOV) domain family of proteins is a well-recognized system for constructing optogenetic devices. Translation of these proteins into efficient cellular sensors is possible by tuning their photochemistry lifetime. However, the bottleneck is the need for more understanding of the relationship between the protein environment and photocycle kinetics. Significantly, the effect of the local environment also modulates the electronic structure of the chromophore, which perturbs the electrostatic and hydrophobic interaction within the binding site. This work highlights the critical factors hidden in the protein network linking with their experimental photocycle kinetics. It also presents an opportunity to quantitatively examine the alternation in chromophore equilibrium geometry and identify details which have substantial implications in designing synthetic constructs with desirable photocycle efficiency.

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“…This photoinhibitory LOV2 insertion strategy is fundamentally different when compared to other LOV2-based optogenetic modulations of activity. In most cases, LOV2 has been employed in an activating scheme in which light absorption results in complete domain dissociation or uncaging of the protein of interest ( 21 – 24 ), or in dimerization of a split protein ( 21 , 25 , 26 ). Domain-level uncaging has more recently been used in a circularly permutated LOV2 system to either activate or inhibit activities in new ways ( 23 ).…”

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confidence: 99%

Allosteric inactivation of an engineered optogenetic GTPase

Jain

Dokholyan

Lee

2023

Proc. Natl. Acad. Sci. U.S.A.

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Optogenetics is a technique for establishing direct spatiotemporal control over molecular function within living cells using light. Light application induces conformational changes within targeted proteins that produce changes in function. One of the applications of optogenetic tools is an allosteric control of proteins via light-sensing domain (LOV2), which allows direct and robust control of protein function. Computational studies supported by cellular imaging demonstrated that application of light allosterically inhibited signaling proteins Vav2, ITSN, and Rac1, but the structural and dynamic basis of such control has yet to be elucidated by experiment. Here, using NMR spectroscopy, we discover principles of action of allosteric control of cell division control protein 42 (CDC42), a small GTPase involved in cell signaling. Both LOV2 and Cdc42 employ flexibility in their function to switch between “dark”/“lit” or active/inactive states, respectively. By conjoining Cdc42 and phototropin1 LOV2 domains into the bi-switchable fusion Cdc42Lov, application of light—or alternatively, mutation in LOV2 to mimic light absorption—allosterically inhibits Cdc42 downstream signaling. The flow and patterning of allosteric transduction in this flexible system are well suited to observation by NMR. Close monitoring of the structural and dynamic properties of dark versus “lit” states of Cdc42Lov revealed lit-induced allosteric perturbations that extend to Cdc42’s downstream effector binding site. Chemical shift perturbations for lit mimic, I539E, have distinct regions of sensitivity, and both the domains are coupled together, leading to bidirectional interdomain signaling. Insights gained from this optoallosteric design will increase our ability to control response sensitivity in future designs.

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Design and engineering of light-sensitive protein switches

Cited by 9 publications

References 66 publications

Advanced near‐infrared light approaches for neuroimaging and neuromodulation

Advanced near‐infrared light approaches for neuroimaging and neuromodulation

Network analysis of chromophore binding site in LOV domain

Allosteric inactivation of an engineered optogenetic GTPase

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