RESUMEN
Many archaea swim by means of archaella. While the archaellum is similar in function to its bacterial counterpart, its structure, composition, and evolution are fundamentally different. Archaella are related to archaeal and bacterial type IV pili. Despite recent advances, our understanding of molecular processes governing archaellum assembly and stability is still incomplete. Here, we determine the structures of Methanococcus archaella by X-ray crystallography and cryo-EM The crystal structure of Methanocaldococcus jannaschii FlaB1 is the first and only crystal structure of any archaellin to date at a resolution of 1.5 Å, which is put into biological context by a cryo-EM reconstruction from Methanococcus maripaludis archaella at 4 Å resolution created with helical single-particle analysis. Our results indicate that the archaellum is predominantly composed of FlaB1. We identify N-linked glycosylation by cryo-EM and mass spectrometry. The crystal structure reveals a highly conserved metal-binding site, which is validated by mass spectrometry and electron energy-loss spectroscopy. We show in vitro that the metal-binding site, which appears to be a widespread property of archaellin, is required for filament integrity.
Asunto(s)
Proteínas Arqueales/metabolismo , Sitios de Unión/fisiología , Metales/metabolismo , Methanococcus/metabolismo , Microscopía por Crioelectrón/métodos , Cristalografía por Rayos X , Citoesqueleto/metabolismo , Glicosilación , Espectrometría de Masas/métodos , Orgánulos/metabolismo , Dominios Proteicos/fisiologíaRESUMEN
It is well known that insulating samples can accumulate electric charges from exposure to an electron beam. How the accumulation of charge affects imaging parameters and sample stability in transmission electron microscopy is poorly understood. To quantify these effects, it is important to know how the charge is distributed within the sample and how it builds up over time. In the present study, we determine the spatial distribution and temporal dynamics of charge accumulation on vitreous ice samples with embedded proteins through a combination of modeling and Fresnel diffraction experiments. Our data reveal a rapid evolution of the charge state on ice upon initial exposure to the electron beam accompanied by charge gradients at the interfaces between ice and carbon films. We demonstrate that ice film movement and charge state variations occur upon electron beam exposure and are dose-rate dependent. Both affect the image defocus through a combination of sample height changes and lensing effects. Our results may be used as a guide to improve sample preparation, data collection, and data processing for imaging of dose-sensitive samples.
RESUMEN
In this work, we have explored the factors which govern mean free path values obtained from off-axis electron holography measurements. Firstly, we explore the topic from a theoretical perspective, and show that the mean amplitude reconstructed from off-axis holograms is due to the coherent portion of the direct, central object-transmitted beam only - it is not affected by the presence or absence of other scattered beams. Secondly, we present a detailed experimental study which compares mean free path values obtained from hologram sideband, centreband, EELS, and TEM measurements as a function of optical collection angle and energy-loss-filtering. These results confirm that the coherent portion of the direct beam defines the mean amplitude, and additionally show that the coherent portion corresponds to the conventional energy-filtered signal (with threshold 5 eV in this work). Finally, we present summary measurements from a selection of different materials, and compare the results against a simple electron scattering model. This study reinforces the claim that the mean amplitude is defined by the energy-filtered direct beam, and confirms that the contributions of elastic and inelastic scattering to the total mean free path are broadly in line with theoretical expectations for these different materials. These results in aggregate indicate that neither experimental collection angles nor enhanced sensitivity to low-loss phonon scattering affect the mean amplitude signal arising from off-axis holography reconstructions, nor the associated mean free path values which are derived from this mean amplitude.
RESUMEN
The structure of core-shell Cu@silica and Ag@Si nanoparticles obtained in one-step through evaporation of elemental precursors by a high-powered electron beam are investigated. The structure of the core and shell of the particles are investigated in order to elucidate their mechanisms of formation and factors affecting the synthesis. It is proposed that the formation of Cu@silica particles is mainly driven by surface tension differences between Cu and Si while the formation of Ag@Si particles is mainly driven by differences in the vapour concentration of the two components.
RESUMEN
The functionalization of ZnO powders with silver nanoparticles (AgNPs) through a novel maple leaf extract mediated biological process was demonstrated. Maple leaf extract was found to be a very effective bioreduction agent for the reduction of silver ions. The reduction rate of Ag(+) into Ag(0) was found to be much faster than other previously reported bioreduction rates and was comparable to the reduction rates obtained through chemical means. The functionalization of ZnO particles with silver nanoparticles through maple leaf extract mediated bioreduction of silver was investigated through UV-visible spectrophotometry, transmission electron microscopy (TEM), and X-ray diffraction analysis. It was found that the ZnO particles were coated with silver nanoparticles 5-20 nm in diameter. The photocatalytic ability of the ZnO particles functionalized with silver nanoparticles was found to be significantly improved compared to the photocatalytic ability of the neat ZnO particles. The silver functionalized ZnO particles reached 90% degradation of the dye an hour before the neat ZnO particles.
Asunto(s)
Acer/química , Nanopartículas del Metal/química , Extractos Vegetales/química , Hojas de la Planta/química , Plata/química , Óxido de Zinc/químicaRESUMEN
There is a need to develop a simple, cheap, and accessible method of treating patients with kidney failure, especially in resource-limited environments such as disaster areas and the developing world due to the inaccessibility of conventional hemodialysis treatments. In this study, we develop a zeolite-polymer composite nanofiber mesh to remove uremic toxins for blood purification. The nanofiber is composed of blood compatible poly(ethylene-co-vinyl alcohol) (EVOH) as the primary matrix polymer and zeolites which are capable of selectively adsorbing uremic toxins such as creatinine. The composite fiber meshes were produced by a cost-effective electrospinning method: electrospinning composite solutions of EVOH and zeolites. Scanning electron microscope (SEM) images revealed that the 7 w/v% EVOH solution produced non-woven fibers with a continuous and smooth morphology. The SEM also showed that over 90% of zeolites in the solution were successfully incorporated into the EVOH nanofibers. Although the barrier properties of the EVOH matrix lowered the creatinine adsorption capacity of the zeolites in the fiber when compared with adsorption to free zeolites, their adsorption capacity was still 67% of the free zeolites. The proposed composite fibers have the potential to be utilized as a new approach to removing nitrogenous waste products from the bloodstream without the requirement of specialized equipment.
RESUMEN
Metal-semiconductor Janus-like nanoparticles with the composition tantalum silicide-silicon (TaSi2/Si) were synthesized for the first time by means of an evaporation method utilizing a high-power electron beam. The composition of the synthesized particles were characterized using high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), selective area electron diffraction (SAED), and energy dispersive X-ray fluorescence (EDX) analysis. The system is compared to previously synthesized core-shell type particles in order to show possible differences responsible for the Janus-like structure forming instead of a core-shell architecture. It is proposed that the production of Janus-like as opposed to core-shell or monophase particles occurs due to the ability of Ta and Si to form compounds and the relative content of Ta and Si atoms in the produced vapour. Based on the results, a potential mechanism of formation for the TaSi2/Si nanoparticles is discussed.