Bright Thermo-resilient and Promiscuous Zombie Protein for Lighting Applications.

Proteins are at the forefront of materials science, with implementations in optical, electrical, and structural materials for transformative and sustainable technologies. Within the biohybrid light-emitting diode (BioHLED) concept, replacing toxic and/or rare photon filters with classical β-barrel fluorescent proteins (FPs) that must withstand irradiation, temperature, oxidation, and dehydration stress, the question if FPs from extremophiles and/or living fossils might be better for lighting applications arises. We addressed this by introducing a thermostable prokaryotic FP, whose inherent promiscuity enables the design of tunable emitting proteins.

Fusing fluorescent proteins and ferritin for protein cage based lighting devices.

Ferritin cages are an effective platform to encapsulate and stabilize a range of active cargoes and present a promising stepping stone towards a wide range of applications. They have been explored for optoelectronic applications in combination with fluorescent proteins towards bio-hybrid light-emitting diodes (Bio-HLEDs) only recently. However, protein integration within the cage or coassembled ferritin cages relies on electrostatic interactions and requires the supercharging of the fluorescent protein that easily compromises functionality and stability.

Elucidating the Supramolecular Interaction of Positively Supercharged Fluorescent Protein with Anionic Phthalocyanines.

Developing bioinspired materials to convert sunlight into electricity efficiently is paramount for sustainable energy production. Fluorescent proteins are promising candidates as photoactive materials due to their high fluorescence quantum yield and absorption extinction coefficients in aqueous media. However, developing artificial bioinspired photosynthetic systems requires a detailed understanding of molecular interactions and energy transfer mechanisms in the required operating conditions.

Bacterial Hybrid Light-Emitting Diodes.

Photon down-converting filters with fluorescent proteins (FPs) are a new frontier in the quest for rare-earth-free and non-toxic color filters for white light-emitting diodes. There are, however, concerns related to the FP purification costs and lack of FP recyclability/reuse. Here, the direct use of bacteria in photon down-converting filters can be of utmost relevance, eliminating purification and allowing in situ production of new FPs.

VARPA: In Silico Additive Screening for Protein-Based Lighting Devices.

Protein optoelectronics is an emerging field facing implementation and stabilization challenges of proteins in harsh non-natural environments, such as dry polymers, inorganic materials, etc., operating at high temperatures/irradiations. In this context, additives promoting structural and functional protein stabilization are paramount to realize highly performing devices.

A Conformationally Stable π-Expanded X-Type Double Helicene Comprising Dihydropyracylene Units with Multistage Redox Behavior.

A π-expanded X-type double [5]helicene comprising dihydropyracylene moieties was synthesized from commercially available acenaphthene. X-ray crystallographic analysis revealed the unique highly twisted structure of the compound resulting in the occurrence of two enantiomers which were separated by chiral HPLC, owing to their high conformational stability. The compound shows strongly bathochromically shifted UV/vis absorption and emission bands with small Stokes shift and considerable photoluminescence quantum yield and circular polarized luminescence response.

Supercharged Fluorescent Protein-Apoferritin Cocrystals for Lighting Applications.

The application of fluorescent proteins (FPs) in optoelectronics is hindered by the need for effective protocols to stabilize them under device preparation and operational conditions. Factors such as high temperatures, irradiation, and organic solvent exposure contribute to the denaturation of FPs, resulting in a low device performance. Herein, we focus on addressing the photoinduced heat generation associated with FP motion and rapid heat transfer.

Superior protein thermophilicity prediction with protein language model embeddings.

Protein thermostability is important in many areas of biotechnology, including enzyme engineering and protein-hybrid optoelectronics. Ever-growing protein databases and information on stability at different temperatures allow the training of machine learning models to predict whether proteins are thermophilic. predictions could reduce costs and accelerate the development process by guiding researchers to more promising candidates.

Genetically Encoded Oligomerization for Protein-Based Lighting Devices.

Implementing proteins in optoelectronics represents a fresh idea toward a sustainable new class of materials with bio-functions that can replace environmentally unfriendly and/or toxic components without losing device performance. However, their native activity (fluorescence, catalysis, and so on) is easily lost under device fabrication/operation as non-native environments (organic solvents, organic/inorganic interfaces, and so on) and severe stress (temperature, irradiation, and so on) are involved. Herein, a gift bow genetically-encoded macro-oligomerization strategy is showcased to promote protein-protein solid interaction enabling i) high versatility with arbitrary proteins, ii) straightforward electrostatic driven control of the macro-oligomer size by ionic strength, and iii) stabilities over months in pure organic solvents and stress scenarios, allowing to integrate them into classical water-free polymer-based materials/components for optoelectronics.

Core-Shell Structured Fluorescent Protein Nanoparticles: New Paradigm Toward Zero-Thermal-Quenching in High-Power Biohybrid Light-Emitting Diodes.

Stable and efficient high-power biohybrid light-emitting diodes (Bio-HLEDs) using fluorescent proteins (FPs) in photon downconverting filters have not been achieved yet, reaching best efficiencies of 130 lm W stable for >5 h. This is related to the rise of the device temperature (70-80 °C) caused by FP-motion and quick heat-transmission in water-based filters, they lead to a strong thermal emission quenching followed by the quick chromophore deactivation via photoinduced H-transfer. To tackle both issues at once, this work shows an elegant concept of a new FP-based nanoparticle, in which the FP core is shielded by a SiO -shell (FP@SiO ) with no loss of the photoluminescence figures-of-merit over years in foreign environments: dry powder at 25 °C (ambient) or constant 50 °C, as well as suspensions in organic solvents.

Alginate beads as a highly versatile test-sample for optoacoustic imaging.

Test-samples are necessary for the development of emerging imaging approaches such as optoacoustics (OA); these can be used to benchmark new labeling agents and instrumentation, or to characterize image analysis algorithms or the inversion required to form the three-dimensional reconstructions. Alginate beads (AlBes) loaded with labeled mammalian or bacterial cells provide a method of creating defined structures of controllable size and photophysical characteristics and are well-suited for both and use. Here we describe a simple and rapid method for efficient and reproducible production of AlBes with specific characteristics and show three example applications with multispectral OA tomography imaging.

Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging.

Reversibly photo-switchable proteins are essential for many super-resolution fluorescence microscopic and optoacoustic imaging methods. However, they have yet to be used as sensors that measure the distribution of specific analytes at the nanoscale or in the tissues of live animals. Here we constructed the prototype of a photo-switchable Ca sensor based on GCaMP5G that can be switched with 405/488-nm light and describe its molecular mechanisms at the structural level, including the importance of the interaction of the core barrel structure of the fluorescent protein with the Ca receptor moiety.

Croconaine-based nanoparticles enable efficient optoacoustic imaging of murine brain tumors.

Contrast enhancement in optoacoustic (photoacoustic) imaging can be achieved with agents that exhibit high absorption cross-sections, high photostability, low quantum yield, low toxicity, and preferential bio-distribution and clearance profiles. Based on advantageous photophysical properties of croconaine dyes, we explored croconaine-based nanoparticles (CR780RGD-NPs) as highly efficient contrast agents for targeted optoacoustic imaging of challenging preclinical tumor targets. Initial characterization of the CR780 dye was followed by modifications using polyethylene glycol and the cancer-targeting c(RGDyC) peptide, resulting in self-assembled ultrasmall particles with long circulation time and active tumor targeting.

Challenging a Preconception: Optoacoustic Spectrum Differs from the Optical Absorption Spectrum of Proteins and Dyes for Molecular Imaging.

Optoacoustic (photoacoustic) imaging has seen marked advances in detection and data analysis, but there is less progress in understanding the photophysics of common optoacoustic contrast agents. This gap blocks the development of novel agents and the accurate analysis and interpretation of multispectral optoacoustic images. To close it, we developed a multimodal laser spectrometer (MLS) to enable the simultaneous measurement of optoacoustic, absorbance, and fluorescence spectra.

Multiplexed whole-animal imaging with reversibly switchable optoacoustic proteins.

We introduce two photochromic proteins for cell-specific in vivo optoacoustic (OA) imaging with signal unmixing in the temporal domain. We show highly sensitive, multiplexed visualization of T lymphocytes, bacteria, and tumors in the mouse body and brain. We developed machine learning-based software for commercial imaging systems for temporal unmixed OA imaging, enabling its routine use in life sciences.

Amplification of photoacoustic effect in bimodal polymer particles by self-quenching of indocyanine green.

A new type of bimodal contrast agent was made that is based on the self-quenching of indocyanine green (ICG) encapsulated in a biocompatible and biodegradable polymer shell. The increasing of a local ICG concentration that is necessary for the obtaining of self-quenching effect was achieved by freezing-induced loading and layer-by-layer assembly. As a result, an efficient photoacoustic(optoacoustic)/fluorescent contrast agent based on composite indocyanine green/polymer particles was successfully prepared and was characterized by fluorescence and photoacoustic(optoacoustic) tomography .

Structure-Based Mutagenesis of Phycobiliprotein smURFP for Optoacoustic Imaging.

Photo- or optoacoustics (OA) imaging is increasingly being used as a non-invasive imaging method that can simultaneously reveal structure and function in deep tissue. However, the most frequent transgenic OA labels are current fluorescent proteins that are not optimized for OA imaging. Thus, they lack OA signal strength, and their absorption maxima are positioned at short wavelengths, thus giving small penetration depths and strong background signals.

Photocontrollable Proteins for Optoacoustic Imaging.

Photocontrollable proteins revolutionized life-science imaging due to their contribution to subdiffraction-resolution optical microscopy. They might have yet another lasting impact on photo- or optoacoustic imaging (OA). OA combines optical contrast with ultrasound detection enabling high-resolution real-time in vivo imaging well-beyond the typical penetration depth of optical methods.

Light-Responsive Size of Self-Assembled Spiropyran-Lysozyme Nanoparticles with Enzymatic Function.

Novel light-responsive nanoassemblies with switchable size and enzymatic activity are built from a protein and a water-soluble spiropyran. Assemblies are created by electrostatic self-assembly in aqueous solution such that the photochromic property of the spiropyran enables light responsiveness. Upon visible light exposure, the aggregate size increases from 200 to 400 nm.

Crystal structure of a biliverdin-bound phycobiliprotein: Interdependence of oligomerization and chromophorylation.

Small, ultra-red fluorescence protein (smURFP) introduces the non-native biliverdin (BV) chromophore to phycobiliproteins (PBPs), allowing them to be used as transgenic labels for in vivo mammalian imaging. Presently, no structural information exists for PBPs bound to the non-native BV chromophore, which limits the further development of smURFP and related proteins as imaging labels or indicators. Here we describe the first crystal structure of a PBP bound to BV.

Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging.

Reversibly switchable fluorescent proteins (rsFPs) have had a revolutionizing effect on life science imaging due to their contribution to sub-diffraction-resolution optical microscopy (nanoscopy). Initial studies showed that their use as labels could also be highly beneficial for emerging photo- or optoacoustic imaging. It could be shown that their use in optoacoustics (i) strongly improves the imaging contrast-to-noise ratio due to modulation and locked-in detection, (ii) facilitates fluence calibration, affording precise measurements of physiological parameters, and finally (iii) could boost spatial resolution following similar concepts as used for nanoscopy.

Revised domain structure of ulvan lyase and characterization of the first ulvan binding domain.

Biomass waste products from green algae have recently been given new life, as these polysaccharides have potential applications in industry, agriculture, and medicine. One such polysaccharide group called ulvans displays many different, potentially useful properties that arise from their structural versatility. Hence, performing structural analyses on ulvan is crucial for future applications.