From biomolecules to biogeochemistry: Exploring the interaction of an indigenous bacterium with gold

Specialised microbial communities colonise the surface of gold particles in soils/sediments, and catalyse gold dissolution and re-precipitation, thereby contributing to the environmental mobility and toxicity of this ‘inert’ precious metal. We assessed the proteomic and physiological response of Serratia proteamaculans, the first metabolically active bacterium enriched and isolated directly from natural gold particles, when exposed to toxic levels of soluble Au3+ (10 μM). The results were compared to a metal-free blank, and to cultures exposed to similarly toxic levels of soluble Cu2+ (0.1 mM); Cu was chosen for comparison because it is closely associated with Au in nature due to similar geochemical properties. A total of 273 proteins were detected from the cells that experienced the oxidative effects of soluble Au, of which 139 (51%) were upregulated with either sole expression (31%) or had synthesis levels greater than the Au-free control (20%). The majority (54%) of upregulated proteins were functionally different from up-regulated proteins in the bacteria-copper treatment. These proteins were related to broad functions involving metabolism and biogenesis, followed by cellular process and signalling, indicating significant specificity for Au. This proteomic study revealed that the bacterium upregulates the synthesis of various proteins related to oxidative stress response (e.g., Monothiol-Glutaredoxin, Thiol Peroxidase, etc.) and cellular damage repair, which leads to the formation of metallic gold nanoparticles less toxic than ionic gold. Therefore, indigenous bacteria may mediate the toxicity of Au through two different yet simultaneous processes: i) repairing cellular components by replenishing damaged proteins and ii) neutralising reactive oxygen species (ROS) by up-regulating the synthesis of antioxidants. By connecting the fields of molecular bacteriology and environmental biogeochemistry, this study is the first step towards the development of biotechnologies based on indigenous bacteria applied to gold bio-recovery and bioremediation of contaminated environments.Santonu K. Sanyal, Tara Pukala, Parul Mittal, Frank Reith, Joël Brugger, Barbara Etschmann, Jeremiah Shuste

Hemin as a generic and potent protein misfolding inhibitor

Abstract not availableYanqin Liu, John A. Carver, Lam H. Ho, Abigail K. Elias, Ian F. Musgrave, Tara L. Pukal

Single-Antenna Temperature- and Humidity-Sounding Microwave Receiver

For humidity and temperature sounding of Earth s atmosphere, a single-antenna/LNA (low-noise amplifier) is needed in place of two separate antennas for the two frequency bands. This results in significant mass and power savings for GeoSTAR that is comprised of hundreds of antennas per frequency channel. Furthermore, spatial anti-aliasing would reduce the number of horns. An anti-aliasing horn antenna will enable focusing the instrument field of view to the hurricane corridor by reducing spatial aliasing, and thus reduce the number of required horns by up to 50 percent. The single antenna/receiver assembly was designed and fabricated by a commercial vendor. The 118 183-GHz horn is based upon a profiled, smooth-wall design, and the OMT (orthomode transducer) on a quad-ridge design. At the input end, the OMT presents four ver y closely spaced ridges [0.0007 in. (18 m)]. The fabricated assembly contains a single horn antenna and low-noise broadband receiver front-end assembly for passive remote sensing of both temperature and humidity profiles in the Earth s atmosphere at 118 and 183 GHz. The wideband feed with dual polarization capability is the first broadband low noise MMIC receiver with the 118 to 183 GHz bandwidth. This technology will significantly reduce PATH/GeoSTAR mass and power while maintaining 90 percent of the measurement capabilities. This is required for a Mission-of-Opportunity on NOAA s GOES-R satellite now being developed, which in turn will make it possible to implement a Decadal-Survey mission for a fraction of the cost and much sooner than would otherwise be possible

Miniature Low-Noise G-Band I-Q Receiver

Weather forecasting, hurricane tracking, and atmospheric science applications depend on humidity sounding of atmosphere. Current instruments provide these measurements from groundbased, airborne, and low Earth orbit (LEO) satellites by measuring radiometric temperature on the flanks of the 183-GHz water vapor line. Miniature, low-noise receivers have been designed that will enable these measurements from a geostationary, thinned array sounder, which is based on hundreds of low-noise receivers that convert the 180-GHz signal directly to baseband in-phase and in-quadrature signals for digitization and correlation. The developed receivers provide a noise temperature of 450 K from 165 to 183 GHz (NF = 4.1 dB), and have a mass of 3 g while consuming 24 mW of power. These are the most sensitive broadband I-Q receivers at this frequency range that operate at room temperature, and are significantly lower in mass and power consumption than previously reported receivers

Norbornene probes for the study of cysteine oxidation

Abstract not availableLisa J. Alcock, Kyle D. Farrell, Mawey T. Akol, Gregory H. Jones, Matthew M. Tierney, Holger B. Kramer, Tara L. Pukala, Gonçalo J.L. Bernardes, Michael V. Perkins, Justin M. Chalke

Fractional deletion of Compound Kushen Injection indicates cytokine signaling pathways are critical for its perturbation of the cell cycle

Published online: 02 October 2019We used computational and experimental biology approaches to identify candidate mechanisms of action of aTraditional Chinese Medicine, Compound Kushen Injection (CKI), in a breast cancer cell line (MDA-MB-231). Because CKI is a complex mixture of plant secondary metabolites, we used a high-performance liquid chromatography (HPLC) fractionation and reconstitution approach to define chemical fractions required for CKI to induce apoptosis. The initial fractionation separated major from minor compounds, and it showed that major compounds accounted for little of the activity of CKI. Furthermore, removal of no single major compound altered the effect of CKI on cell viability and apoptosis. However, simultaneous removal of two major compounds identified oxymatrine and oxysophocarpine as critical with respect to CKI activity. Transcriptome analysis was used to correlate compound removal with gene expression and phenotype data. Many compounds in CKI are required to trigger apoptosis but significant modulation of its activity is conferred by a small number of compounds. In conclusion, CKI may be typical of many plant based extracts that contain many compounds in that no single compound is responsible for all of the bioactivity of the mixture and that many compounds interact in a complex fashion to influence a network containing many targets.T. N . Aung, S. Nourmohammadi, Z. Qu, Y. Harata-Lee, J. Cui, H. Y. Shen, A. J. Yool, T. Pukala, Hong Du, R. D. Kortschak, W. Wei, D. L. Adelso

Norbornene probes for the study of cysteine oxidation

© 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license http://creativecommons.org/licenses/by/4.0/).Cysteine residues on proteins can react with cellular oxidants such as hydrogen peroxide. While this process is important for scavenging excess reactive oxygen species, the products of this oxidation may also mediate cell signalling. To understand the role of cysteine oxidation in biology, selective probes are required to detect and quantify its occurrence. Cysteine oxidation products such as sulfenic acids are sometimes unstable and therefore short-lived. If such cysteine derivatives are to be analysed, rapid reaction with the probe is required. Here we introduce norbornene derivatives as probes for cysteine oxidation, and demonstrate their ability to trap sulfenic acids. The synthesis of norbornene derivatives containing alkyne or biotin affinity tags are also reported to facilitate the use of these probes in chemical biology and proteomics

Norbornene probes for the study of cysteine oxidation

Cysteine residues on proteins can react with cellular oxidants such as hydrogen peroxide. While this process is important for scavenging excess reactive oxygen species, the products of this oxidation may also mediate cell signalling. To understand the role of cysteine oxidation in biology, selective probes are required to detect and quantify its occurence. Cysteine oxidation products such as sulfenic acids are sometimes unstable and therefore short-lived. If such cysteine derivatives are to be analysed, rapid reaction with the probe is required. Here we introduce norbornene derivatives as probes for cysteine oxidation, and demonstrate their ability to trap sulfenic acids. The synthesis of norbornene derivatives containing alkyne or biotin affinity tags are also reported to facilitate the use of these probes in chemical biology and proteomics

Advanced resistance studies identify two discrete mechanisms in staphylococcus aureus to overcome antibacterial compounds that target biotin protein ligase

Biotin protein ligase (BPL) inhibitors are a novel class of antibacterial that target clinically important methicillin-resistant Staphylococcus aureus (S. aureus). In S. aureus, BPL is a bifunctional protein responsible for enzymatic biotinylation of two biotin-dependent enzymes, as well as serving as a transcriptional repressor that controls biotin synthesis and import. In this report, we investigate the mechanisms of action and resistance for a potent anti-BPL, an antibacterial compound, biotinyl-acylsulfamide adenosine (BASA). We show that BASA acts by both inhibiting the enzymatic activity of BPL in vitro, as well as functioning as a transcription co-repressor. A low spontaneous resistance rate was measured for the compound (<10-9) and whole-genome sequencing of strains evolved during serial passaging in the presence of BASA identified two discrete resistance mechanisms. In the first, deletion of the biotin-dependent enzyme pyruvate carboxylase is proposed to prioritize the utilization of bioavailable biotin for the essential enzyme acetyl-CoA carboxylase. In the second, a D200E missense mutation in BPL reduced DNA binding in vitro and transcriptional repression in vivo. We propose that this second resistance mechanism promotes bioavailability of biotin by derepressing its synthesis and import, such that free biotin may outcompete the inhibitor for binding BPL. This study provides new insights into the molecular mechanisms governing antibacterial activity and resistance of BPL inhibitors in S. aureus.Andrew J. Hayes, Jiulia Satiaputra, Louise M. Sternicki, Ashleigh S. Paparella, Zikai Feng, Kwang J. Lee ... et al