Protein interaction networks and protein compartmentalization underlie all signaling and regulatory processes in cells. Enzyme-catalyzed proximity labeling (PL) has emerged as a new approach to study the spatial and interaction characteristics of proteins in living cells. However, current PL methods require over 18 h of labeling time or utilize chemicals with limited cell permeability or high toxicity. We used yeast display-based directed evolution to engineer two promiscuous mutants of biotin ligase, TurboID and miniTurbo, which catalyze PL with much greater efficiency than BioID or BioID2, and enable 10-min PL in cells with non-toxic and easily deliverable biotin. Furthermore, TurboID extends biotin-based PL to flies and worms.

Efficient proximity labeling in living cells and organisms with TurboID.

and Perspectives Sophie Carenco,†,‡,§,∥,⊥ David Portehault,*,†,‡,§ Ced́ric Boissier̀e,†,‡,§ Nicolas Meźailles, and Cleḿent Sanchez*,†,‡,§ †Chimie de la Matier̀e Condenseé de Paris, UPMC Univ Paris 06, UMR 7574, Colleg̀e de France, 11 Place Marcelin Berthelot, 75231 Paris Cedex 05, France ‡Chimie de la Matier̀e Condenseé de Paris, CNRS, UMR 77574, Colleg̀e de France, 11 Place Marcellin Berthelot, 75231 Paris Cedex 05, France Chimie de la Matier̀e Condenseé de Paris, Colleg̀e de France, 11 Place Marcellin Berthelot, 75231 Paris Cedex 05, France Laboratory Heteroelements and Coordination, Chemistry Department, Ecole Polytechnique, CNRS-UMR 7653, Palaiseau, France

Nanoscaled metal borides and phosphides: recent developments and perspectives

Progress in corneal wound healing

Inspired by nanoscience and nanoengineering, numerous nanostructured materials developed by multidisciplinary approaches exhibit excellent photoelectronic properties ranging from ultraviolet to terahertz frequencies. As a new class of building block, nanoscale elements in terms of quantum dots, nanowires, and nanolayers can be used for fabricating photodetectors with high performance. Moreover, in conjunction with traditional photodetectors, they exhibit appealing performance for practical applications including high density of integration, high sensitivity, fast response, and multifunction. Therefore, with the perspective of photodetectors constructed by diverse low-dimensional nanostructured materials, recent advances in nanoscale photodetectors are discussed here; meanwhile, challenges and promising future directions in this research field are proposed.

Nanostructured Photodetectors: From Ultraviolet to Terahertz.

Electromagnetic absorbers have drawn increasing attention in many areas. A series of plasmonic and metamaterial structures can work as efficient narrowband absorbers due to the excitation of plasmonic or photonic resonances, providing a great potential for applications in designing selective thermal emitters, biosensing, etc. In other applications such as solar-energy harvesting and photonic detection, the bandwidth of light absorbers is required to be quite broad. Under such a background, a variety of mechanisms of broadband/multiband absorption have been proposed, such as mixing multiple resonances together, exciting phase resonances, slowing down light by anisotropic metamaterials, employing high loss materials and so on.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/lpor.201400026

Plasmonic and metamaterial structures as electromagnetic absorbers

Toxoplasma gondii is an obligate intracellular parasite that invades host cells and replicates within a unique parasitophorous vacuole. To maintain this intracellular niche, the parasite secretes an array of dense granule proteins (GRAs) into the nascent parasitophorous vacuole. These GRAs are believed to play key roles in vacuolar remodeling, nutrient uptake, and immune evasion while the parasite is replicating within the host cell. Despite the central role of GRAs in the Toxoplasma life cycle, only a subset of these proteins have been identified, and many of their roles have not been fully elucidated. In this report, we utilize the promiscuous biotin ligase BirA* to biotinylate GRA proteins secreted into the vacuole and then identify those proteins by affinity purification and mass spectrometry. Using GRA-BirA* fusion proteins as bait, we have identified a large number of known and candidate GRAs and verified localization of 13 novel GRA proteins by endogenous gene tagging. We proceeded to functionally characterize three related GRAs from this group (GRA38, GRA39, and GRA40) by gene knockout. While Δgra38 and Δgra40 parasites showed no altered phenotype, disruption of GRA39 results in slow-growing parasites that contain striking lipid deposits in the parasitophorous vacuole, suggesting a role in lipid regulation that is important for parasite growth. In addition, parasites lacking GRA39 showed dramatically reduced virulence and a lower tissue cyst burden in vivo. Together, the findings from this work reveal a partial vacuolar proteome of T. gondii and identify a novel GRA that plays a key role in parasite replication and pathogenesis. IMPORTANCE Most intracellular pathogens reside inside a membrane-bound vacuole within their host cell that is extensively modified by the pathogen to optimize intracellular growth and avoid host defenses. In Toxoplasma, this vacuole is modified by a host of secretory GRA proteins, many of which remain unidentified. Here we demonstrate that in vivo biotinylation of proximal and interacting proteins using the promiscuous biotin ligase BirA* is a powerful approach to rapidly identify vacuolar GRA proteins. We further demonstrate that one factor identified by this approach, GRA39, plays an important role in the ability of the parasite to replicate within its host cell and cause disease.

/pdf/in-vivo-biotinylation-of-the-toxoplasma-parasitophorous-33b4fpjo1b.pdf

In Vivo Biotinylation of the Toxoplasma Parasitophorous Vacuole Reveals Novel Dense Granule Proteins Important for Parasite Growth and Pathogenesis

The directed growth of III–V nanopillars is used to demonstrate bottom-up photonic crystal lasers. Simultaneous formation of both the photonic band gap and active gain region is achieved via catalyst-free selective-area metal–organic chemical vapor deposition on masked GaAs substrates. The nanopillars implement a GaAs/InGaAs/GaAs axial double heterostructure for accurate, arbitrary placement of gain within the cavity and lateral InGaP shells to reduce surface recombination. The lasers operate single-mode at room temperature with low threshold peak power density of ∼625 W/cm2. Cavity resonance and lasing wavelength is lithographically defined by controlling pillar pitch and diameter to vary from 960 to 989 nm. We envision this bottom-up approach to pillar-based devices as a new platform for photonic systems integration.

https://authors.library.caltech.edu/28791/2/nl2030163_si_001.pdf

Bottom-up Photonic Crystal Lasers

We demonstrate nanopillar- (NP) based plasmon-enhanced photodetectors (NP-PEPDs) operating in the near-infrared spectral regime. A novel fabrication technique produces subwavelength elongated nanoholes in a metal surface self-aligned to patterned NP arrays that acts as a 2D plasmonic crystal. Surface plasmon Polariton Bloch waves (SPP-BWs) are excited by the metal nanohole array resulting in electric field intensity “hot spots” in the NP. The NP periodicity determines the peak responsivity wavelength while the nanohole asymmetry produces polarization-dependent coupling of the SPP-BW modes. Resulting photodetectors have 0.28 A/W responsivity peaked at 1100 nm at a reverse bias of −5 V. Designs for further increasing the optical coupling efficiency into the nanopillar are explored. This technology has potential applications for plasmonically enhanced focal plane arrays and plasmonic photovoltaics.

/pdf/surface-plasmon-enhanced-nanopillar-photodetectors-2yh3kiehg5.pdf

Surface plasmon-enhanced nanopillar photodetectors.

In this work, we study hybrid solar cells based on poly(3-hexylthiophene)-coated GaAs nanopillars grown on a patterned GaAs substrate using selective-area metal organic chemical vapor deposition The hybrid solar cells show extremely low leakage currents (I≅45 nA @−1V) under dark conditions and an open circuit voltage, short circuit current density, and fill factor of 02 V, 87 mA/cm2, and 32%, respectively, giving a power conversion efficiency of η=06% under AM 15 G illumination Surface passivation of the GaAs results in further improvement, yielding η=144% under AM 15 G illumination External quantum efficiency measurements of these polymer/inorganic solar cells are also presented

/pdf/hybrid-conjugated-polymer-solar-cells-using-patterned-gaas-1bl4d5clem.pdf

Hybrid conjugated polymer solar cells using patterned GaAs nanopillars

The Toxoplasma inner membrane complex (IMC) is a specialized organelle underlying the parasite's plasma membrane that consists of flattened rectangular membrane sacs that are sutured together and positioned atop a supportive cytoskeleton. We have previously identified a novel class of proteins localizing to the transverse and longitudinal sutures of the IMC, which we named IMC sutures components (ISCs). Here, we have used proximity-dependent biotin identification at the sutures to better define the composition of this IMC subcompartment. Using ISC4 as bait, we demonstrate biotin-dependent labeling of the sutures and have uncovered two new ISCs. We also identified five new proteins that exclusively localize to the transverse sutures that we named transverse sutures components (TSCs), demonstrating that components of the IMC sutures consist of two groups: those that localize to the transverse and longitudinal sutures (ISCs) and those residing only in the transverse sutures (TSCs). In addition, we functionally analyze the ISC protein ISC3 and demonstrate that ISC3-null parasites have morphological defects and reduced fitness in vitro. Most importantly, Δisc3 parasites exhibit a complete loss of virulence in vivo. These studies expand the known composition of the IMC sutures and highlight the contribution of ISCs to the ability of the parasite to proliferate and cause disease.

/pdf/novel-insights-into-the-composition-and-function-of-the-2l7m6xlxb8.pdf

Andrew Lin

Papers

In Vivo Biotinylation of the Toxoplasma Parasitophorous Vacuole Reveals Novel Dense Granule Proteins Important for Parasite Growth and Pathogenesis

Bottom-up Photonic Crystal Lasers

Surface plasmon-enhanced nanopillar photodetectors.

Hybrid conjugated polymer solar cells using patterned GaAs nanopillars

Novel insights into the composition and function of the Toxoplasma IMC sutures.