Heterogeneous nucleation is required for crystallization of the ZnuA domain of pneumococcal AdcA

Zn2+ is an essential nutrient for all known forms of life. In the major human pathogen Streptococcus pneumoniae, the acquisition of Zn2+ is facilitated by two Zn2+-specific solute-binding proteins: AdcA and AdcAII. To date, there has been a paucity of structural information on AdcA, which has hindered a deeper understanding of the mechanism underlying pneumococcal Zn2+ acquisition. Native AdcA consists of two domains: an N-terminal ZnuA domain and a C-terminal ZinT domain. In this study, the ZnuA domain of AdcA was crystallized. The initial crystals of the ZnuA-domain protein were obtained using dried seaweed as a heterogeneous nucleating agent. No crystals were obtained in the absence of the heterogeneous nucleating agent. These initial crystals were subsequently used as seeds to produce diffraction-quality crystals. The crystals diffracted to 2.03 angstrom resolution and had the symmetry of space group P1. This study demonstrates the utility of heterogeneous nucleation. The solution of the crystal structures will lead to further understanding of Zn2+ acquisition by S. pneumoniae

Face Model Compression by Distilling Knowledge from Neurons

The recent advanced face recognition systems werebuilt on large Deep Neural Networks (DNNs) or theirensembles, which have millions of parameters. However, the expensive computation of DNNs make theirdeployment difficult on mobile and embedded devices. This work addresses model compression for face recognition,where the learned knowledge of a large teachernetwork or its ensemble is utilized as supervisionto train a compact student network. Unlike previousworks that represent the knowledge by the soften labelprobabilities, which are difficult to fit, we represent theknowledge by using the neurons at the higher hiddenlayer, which preserve as much information as the label probabilities, but are more compact. By leveragingthe essential characteristics (domain knowledge) of thelearned face representation, a neuron selection methodis proposed to choose neurons that are most relevant toface recognition. Using the selected neurons as supervisionto mimic the single networks of DeepID2+ andDeepID3, which are the state-of-the-art face recognition systems, a compact student with simple network structure achieves better verification accuracy on LFW than its teachers, respectively. When using an ensemble of DeepID2+ as teacher, a mimicked student is able to outperform it and achieves 51.6 times compression ratio and 90 times speed-up in inference, making this cumbersome model applicable on portable devices

A Trap-Door Mechanism for Zinc Acquisition by Streptococcus pneumoniae AdcA

Zinc is an essential element in all domains of life. Nonetheless, how pro-karyotes achieve selective acquisition of zinc from the extracellular environment remains poorly understood. Here, we elucidate a novel mechanism for zinc-binding in AdcA, a solute-binding protein of Streptococcus pneumoniae. Crystal structure analyses reveal the two-domain organization of the protein and show that only the N-terminal domain (AdcAN) is necessary for zinc import. Zinc binding induces only minor changes in the global protein conformation of AdcA and stabilizes a highly mobile loop within the AdcAN domain. This loop region, which is conserved in zinc-specific solute-binding proteins, facilitates closure of the AdcAN binding site and is crucial for zinc acquisition. Collectively, these findings elucidate the structural and functional basis of selective zinc uptake in prokaryotes. IMPORTANCE Zinc is an essential nutrient for the virulence of bacterial pathogens such as Streptococcus pneumoniae. Many Gram-positive bacteria use a two-do-main lipoprotein for zinc acquisition, but how this class of metal-recruiting proteins acquire zinc and interact with the uptake machinery has remained poorly defined. We report the first structure of a two-domain lipoprotein, AdcA from S. pneumoniae, and use computational, spectroscopic, and microbiological approaches to provide new insights into the functional basis of zinc recruitment. Our findings reveal that AdcA employs a novel mechanism for zinc binding that we have termed the “trap-door” mechanism, and we show how the static metal-binding site of the protein, which confers its selectivity for zinc ions, is combined with a dynamic surface element to facilitate zinc recruitment and import into the bacte-rium. Together, these findings expand our understanding of how bacteria acquire zinc from the environment and provide a foundation for inhibiting this process, through antimicrobial targeting of the dynamic structural elements to block bacterial zinc scavenging.</p

Long-Term Stability of Different Kinds of Gas Nanobubbles in Deionized and Salt Water

Nanobubbles have many potential applications depending on their types. The long-term stability of different gas nanobubbles is necessary to be studied considering their applications. In the present study, five kinds of nanobubbles (N2, O2, Ar + 8%H2, air and CO2) in deionized water and a salt aqueous solution were prepared by the hydrodynamic cavitation method. The mean size and zeta potential of the nanobubbles were measured by a light scattering system, while the pH and Eh of the nanobubble suspensions were measured as a function of time. The nanobubble stability was predicted and discussed by the total potential energies between two bubbles by the extended Derjaguin–Landau–Verwey–Overbeek (DLVO) theory. The nanobubbles, except CO2, in deionized water showed a long-term stability for 60 days, while they were not stable in the 1 mM (milli mol/L) salt aqueous solution. During the 60 days, the bubble size gradually increased and decreased in deionized water. This size change was discussed by the Ostwald ripening effect coupled with the bubble interaction evaluated by the extended DLVO theory. On the other hand, CO2 nanobubbles in deionized water were not stable and disappeared after 5 days, while the CO2 nanobubbles in 1 mM of NaCl and CaCl2 aqueous solution became stable for 2 weeks. The floating and disappearing phenomena of nanobubbles were estimated and discussed by calculating the relationship between the terminal velocity of the floating bubble and bubble size.Validerad;2021;Nivå 2;2021-04-16 (alebob);Finansiär: Natural Science Foundation of China (NSFC 21976039)</p

Fabrication of Chiral 3D Microstructure Using Tightly Focused Multiramp Helico-Conical Optical Beams

Beams with optical vortices are widely used in various fields, including optical communication, optical manipulation and trapping, and, especially in recent years, in the processing of nanoscale structures. However, circular vortex beams are difficult to use for the processing of chiral micro and nanostructures. This paper introduces a multiramp helical&ndash;conical beam that can produce a three-dimensional spiral light field in a tightly focused system. Using this spiral light beam and the two-photon direct writing technique, micro&ndash;nano structures with chiral characteristics in space can be directly written under a single exposure. The fabrication efficiency is more than 20 times higher than the conventional point-by-point writing strategy. The tightly focused properties of the light field were utilized to analyze the field-dependent properties of the micro&ndash;nano structure, such as the number of multiramp mixed screw-edge dislocations. Our results enrich the means of two-photon polymerization technology and provide a simple and stable way for the micromachining of chiral microstructures, which may have a wide range of applications in optical tweezers, optical communications, and metasurfaces

The structure and metal binding properties of Chlamydia trachomatis YtgA

The obligate intracellular pathogen is a globally significant cause of sexually transmitted bacterial infections and the leading etiological agent of preventable blindness. The first-row transition metal iron (Fe) plays critical roles in chlamydial cell biology, and acquisition of this nutrient is essential for the survival and virulence of the pathogen. Nevertheless, how acquires Fe from host cells is not well understood, since it lacks genes encoding known siderophore biosynthetic pathways, receptors for host Fe storage proteins, and the Fe acquisition machinery common to many bacteria. Recent studies have suggested that directly acquires host Fe via the ATP-binding cassette permease YtgABCD. Here, we characterized YtgA, the periplasmic solute binding protein component of the transport pathway, which has been implicated in scavenging Fe(III) ions. The structure of Fe(III)-bound YtgA was determined at 2.0-Å resolution with the bound ion coordinated via a novel geometry (3 Ns, 2 Os [3N2O]). This unusual coordination suggested a highly plastic metal binding site in YtgA capable of interacting with other cations. Biochemical analyses showed that the metal binding site of YtgA was not restricted to interaction with only Fe(III) ions but could bind all transition metal ions examined. However, only Mn(II), Fe(II), and Ni(II) ions bound reversibly to YtgA, with Fe being the most abundant cellular transition metal in Collectively, these findings show that YtgA is the metal-recruiting component of the YtgABCD permease and is most likely involved in the acquisition of Fe(II) and Mn(II) from host cells. is the most common bacterial sexually transmitted infection in developed countries, with an estimated global prevalence of 4.2% in the 15- to 49-year age group. Although infection is asymptomatic in more than 80% of infected women, about 10% of cases result in serious disease. Infection by is dependent on the ability to acquire essential nutrients, such as the transition metal iron, from host cells. In this study, we show that iron is the most abundant transition metal in and report the structural and biochemical properties of the iron-recruiting protein YtgA. Knowledge of the high-resolution structure of YtgA will provide a platform for future structure-based antimicrobial design approaches