Intercellular protein-protein interactions (PPIs) enable communication between cells in diverse biological processes, including cell proliferation, immune responses, infection, and synaptic ...transmission, but they are challenging to visualize because existing techniques have insufficient sensitivity and/or specificity. Here we report a split horseradish peroxidase (sHRP) as a sensitive and specific tool for the detection of intercellular PPIs. The two sHRP fragments, engineered through screening of 17 cut sites in HRP followed by directed evolution, reconstitute into an active form when driven together by an intercellular PPI, producing bright fluorescence or contrast for electron microscopy. Fusing the sHRP fragments to the proteins neurexin (NRX) and neuroligin (NLG), which bind each other across the synaptic cleft, enabled sensitive visualization of synapses between specific sets of neurons, including two classes of synapses in the mouse visual system. sHRP should be widely applicable to studying mechanisms of communication between a variety of cell types.
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IJS, NUK, SBMB, UL, UM, UPUK
ABSTRACT
Streams of stars from captured dwarf galaxies and dissolved globular clusters are identifiable through the similarity of their orbital parameters, a fact that remains true long after the ...streams have dispersed spatially. We calculate the integrals of motion for 31 234 stars, to a distance of 4 kpc from the Sun, which have full and accurate 6D phase space positions in the Gaia DR2 catalogue. We then apply a novel combination of data mining, numerical, and statistical techniques to search for stellar streams. This process returns five high confidence streams (including one which was previously undiscovered), all of which display tight clustering in the integral of motion space. Colour–magnitude diagrams indicate that these streams are relatively simple, old, metal-poor populations. One of these resolved streams shares very similar kinematics and metallicity characteristics with the Gaia-Enceladus dwarf galaxy remnant, but with a slightly younger age. The success of this project demonstrates the usefulness of data mining techniques in exploring large data sets.
We present high-resolution Magellan/MIKE spectroscopy of 42 red giant stars in seven stellar streams confirmed by the Southern Stellar Stream Spectroscopic Survey (S5): ATLAS, Aliqa Uma, Chenab, ...Elqui, Indus, Jhelum, and Phoenix. Abundances of 30 elements have been derived from over 10,000 individual line measurements or upper limits using photometric stellar parameters and a standard LTE analysis. This is currently the most extensive set of element abundances for stars in stellar streams. Three streams (ATLAS, Aliqa Uma, and Phoenix) are disrupted metal-poor globular clusters, although only weak evidence is seen for the light-element anticorrelations commonly observed in globular clusters. Four streams (Chenab, Elqui, Indus, and Jhelum) are disrupted dwarf galaxies, and their stars display abundance signatures that suggest progenitors with stellar masses ranging from 106 to 107 M . Extensive description is provided for the analysis methods, including the derivation of a new method for including the effect of stellar parameter correlations on each star's abundance and uncertainty. This paper includes data gathered with the 6.5 m Magellan Telescopes located at Las Campanas Observatory, Chile.
ABSTRACT
In order to accurately determine stellar properties, knowledge of the effective temperature of stars is vital. We implement Gaia and 2MASS photometry in the InfraRed Flux Method and apply it ...to over 360 000 stars across different evolutionary stages in the GALAH DR3 survey. We derive colour-effective temperature relations that take into account the effect of metallicity and surface gravity over the range $4000\, \rm {K}\lesssim T_{\rm {eff}}\lesssim 8000\, \rm {K}$, from very metal-poor stars to supersolar metallicities. The internal uncertainty of these calibrations is of order 40–80 K depending on the colour combination used. Comparison against solar-twins, Gaia benchmark stars, and the latest interferometric measurements validates the precision and accuracy of these calibrations from F to early M spectral types. We assess the impact of various sources of uncertainties, including the assumed extinction law, and provide guidelines to use our relations. Robust solar colours are also derived.
The widespread deployment of carbon capture and sequestration as a climate change mitigation strategy could be facilitated by the development of more energy-efficient adsorbents. Diamine-appended ...metal-organic frameworks of the type diamine-M
(dobpdc) (M = Mg, Mn, Fe, Co, Ni, Zn; dobpdc
= 4,4'-dioxidobiphenyl-3,3'-dicarboxylate) have shown promise for carbon-capture applications, although questions remain regarding the molecular mechanisms of CO
uptake in these materials. Here we leverage the crystallinity and tunability of this class of frameworks to perform a comprehensive study of CO
chemisorption. Using multinuclear nuclear magnetic resonance (NMR) spectroscopy experiments and van-der-Waals-corrected density functional theory (DFT) calculations for 13 diamine-M
(dobpdc) variants, we demonstrate that the canonical CO
chemisorption products, ammonium carbamate chains and carbamic acid pairs, can be readily distinguished and that ammonium carbamate chain formation dominates for diamine-Mg
(dobpdc) materials. In addition, we elucidate a new chemisorption mechanism in the material dmpn-Mg
(dobpdc) (dmpn = 2,2-dimethyl-1,3-diaminopropane), which involves the formation of a 1:1 mixture of ammonium carbamate and carbamic acid and accounts for the unusual adsorption properties of this material. Finally, we show that the presence of water plays an important role in directing the mechanisms for CO
uptake in diamine-M
(dobpdc) materials. Overall, our combined NMR and DFT approach enables a thorough depiction and understanding of CO
adsorption within diamine-M
(dobpdc) compounds, which may aid similar studies in other amine-functionalized adsorbents in the future.
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IJS, KILJ, NUK, PNG, UL, UM
ABSTRACT
Gaia DR2 has revealed new small-scale and large-scale patterns in the phase-space distribution of stars in the Milky Way. In cylindrical Galactic coordinates $(R,\phi ,z)$, ridge-like ...structures can be seen in the $(R,V_\phi)$ plane and asymmetric arch-like structures in the $(V_R,V_\phi)$ plane. We show that the ridges are also clearly present when the third dimension of the $(R,V_\phi)$ plane is represented by $\langle z \rangle$, $\langle V_z \rangle$, $\langle V_R \rangle$, $\langle$Fe/H$\rangle$, and $\langle \alpha /{\rm Fe}\rangle$. The maps suggest that stars along the ridges lie preferentially close to the Galactic mid-plane ($|z|\lt 0.2$ kpc), and have metallicity and $\alpha$ elemental abundance similar to that of the Sun. We show that phase mixing of disrupting spiral arms can generate both the ridges and the arches. It also generates discrete groupings in orbital energy – the ridges and arches are simply surfaces of constant energy. We identify eight distinct ridges in the Gaia DR2 data: six of them have constant energy while two have constant angular momentum. Given that the signature is strongest for stars close to the plane, the presence of ridges in $\langle z \rangle$ and $\langle V_z \rangle$ suggests a coupling between planar and vertical directions. We demonstrate, using N-body simulations that such coupling can be generated both in isolated discs and in discs perturbed by an orbiting satellite like the Sagittarius dwarf galaxy.
Proximity labeling (PL) has emerged as a powerful approach to elucidate proteomes within a defined radius around a protein of interest (POI). In PL, a catalyst is attached to the POI and tags nearby ...endogenous proteins, which are then isolated by affinity purification and identified by mass spectrometry. Although existing PL methods have yielded numerous biological insights, proteomes with greater spatial resolution could be obtained if PL catalysts could be activated at more specific subcellular locations, such as sites where both the POI and a chemical stimulus are present or sites of protein–protein interactions (PPIs). Here, we report DNA-based switchable PL catalysts that are attached to a POI and become activated only when a secondary molecular trigger is present. The DNA catalysts consist of a photocatalyst and a spectral quencher tethered to a DNA oligomer. They are catalytically inactive by default but undergo a conformational change in response to a specific molecular trigger, thus activating PL. We designed a system in which the DNA catalyst becomes activated on living mammalian cells specifically at sites of Her2–Her3 heterodimers and c-Met homodimers, PPIs known to increase the invasion and growth of certain cancers. While this study employs a Ru(bpy)3-type complex for tagging proteins with biotin phenol, the switchable DNA catalyst design is compatible with diverse synthetic PL photocatalysts. Furthermore, the switchable DNA PL catalysts can be constructed from conformation-switching DNA aptamers that respond to small molecules, ions, and proteins, opening future opportunities for PL in highly specific subcellular locations.
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IJS, KILJ, NUK, PNG, UL, UM
APEX is an engineered peroxidase that catalyzes the oxidation of a wide range of substrates, facilitating its use in a variety of applications from subcellular staining for electron microscopy to ...proximity biotinylation for spatial proteomics and transcriptomics. To further advance the capabilities of APEX, we used directed evolution to engineer a split APEX tool (sAPEX). A total of 20 rounds of fluorescence activated cell sorting (FACS)-based selections from yeast-displayed fragment libraries, using 3 different surface display configurations, produced a 200-amino-acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino-acid C-terminal fragment called “EX”. AP and EX fragments were each inactive on their own but were reconstituted to give peroxidase activity when driven together by a molecular interaction. We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within a non-coding RNA scaffold, and at mitochondria–endoplasmic reticulum contact sites.
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IJS, KILJ, NUK, PNG, UL, UM, UPUK
Globular clusters are some of the oldest bound stellar structures observed in the Universe
. They are ubiquitous in large galaxies and are believed to trace intense star-formation events and the ...hierarchical build-up of structure
. Observations of globular clusters in the Milky Way, and a wide variety of other galaxies, have found evidence for a 'metallicity floor', whereby no globular clusters are found with chemical (metal) abundances below approximately 0.3 to 0.4 per cent of that of the Sun
. The existence of this metallicity floor may reflect a minimum mass and a maximum redshift for surviving globular clusters to form-both critical components for understanding the build-up of mass in the Universe
. Here we report measurements from the Southern Stellar Streams Spectroscopic Survey of the spatially thin, dynamically cold Phoenix stellar stream in the halo of the Milky Way. The properties of the Phoenix stream are consistent with it being the tidally disrupted remains of a globular cluster. However, its metal abundance (Fe/H = -2.7) is substantially below the empirical metallicity floor. The Phoenix stream thus represents the debris of the most metal-poor globular clusters discovered so far, and its progenitor is distinct from the present-day globular cluster population in the local Universe. Its existence implies that globular clusters below the metallicity floor have probably existed, but were destroyed during Galactic evolution.
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FZAB, GEOZS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
ABSTRACT
We report the discovery of the only very nitrogen-enhanced metal-poor star known in a Galactic globular cluster. This star, in the very metal-poor cluster ESO280−SC06 , has ...$\textrm{N}/\textrm{Fe}\, \gt +2.5$, while the other stars in the cluster show no obvious enhancement in nitrogen. Around 80 NEMP stars are known in the field, and their abundance patterns are believed to reflect mass transfer from a binary companion in the asymptotic giant branch phase. The dense environment of globular clusters is detrimental to the long term survival of binary systems, resulting in a low observed binary fraction among red giants and the near absence of NEMP stars. We also identify the first known horizontal branch members of ESO280−SC06 , which allow for a much better constraint on its distance. We calculate an updated orbit for the cluster based on our revised distance of 20.6 ± 0.5 kpc, and find no significant change to its orbital properties.
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