Molecular characterization of an anion pump. The ArsB protein is the membrane anchor for the ArsA protein

R-factor mediated bacterial resistance to arsenical salts occurs by active extrusion of the toxic oxyanions from cells of gram negative bacteria. The ars operon of the conjugative plasmid R773 encodes an anion pump. The pump has two polypeptide components. The catalytic subunit, the ArsA protein, is an oxyanion-stimulated ATPase. The membrane component, the ArsB protein, has been localized in the inner membrane of Escherichia coli. The ArsA and ArsB proteins have been postulated to form a membrane complex which functions as an anion-translocating ATPase. In this study evidence is presented showing that expression of the arsB gene is required to anchor the ArsA protein to the inner membrane. Binding studies with purified ArsA to membranes with and without the arsB gene product confirm this requirement. Membranes of uncA mutants containing both the ArsA and ArsB proteins exhibit arsenite(antimonite)-stimulated ATPase activity. These results support the model in which the ArsA protein is the catalytic energy transducing component of the anion pump, whereas the integral membrane ArsB protein serves as both the anion channel and membrane binding site for the ArsA protein

Membrane topology of the ArsB protein, the membrane subunit of an anion-translocating ATPase

The ars operon of the conjugative R-factor R773 encodes an oxyanion pump that catalyzes extrusion of arsenicals from cells of Escherichia coli. The oxyanion translocation ATPase is composed of two polypeptides, the catalytic ArsA protein and the intrinsic membrane protein, ArsB. The topology of regions of the ArsB protein in the inner membrane was determined using a variety of gene fusions. Random gene fusions with lacZ and phoA were generated using transposon mutagenesis. A series of gene fusions with blaM were constructed in vitro using a beta-lactamase fusion vector. To localize individual segments of the ArsB protein, a ternary fusion method was developed, where portions of the arsB gene were inserted in-frame between the coding regions for two heterologous proteins, in this case a portion of a newly identified arsD gene and the blaM sequence encoding the mature beta-lactamase. The location of a periplasmic loop was determined from V8 protease digestion of an ArsA-ArsB chimera. From analysis of data from 26 fusions, a topological model of the ArsB protein with 12 membrane-spanning regions is proposed

New ⁴⁰Ar/³⁹Ar dating of the Grande Ronde lavas, Columbia River Basalts, USA: Implications for duration of flood basalt eruption episodes

Grande Ronde Basalt (GRB) lavas represent the most voluminous eruptive pulse of the Columbia River-Snake River-Yellowstone hotspot volcanism. With an estimated eruptive volume of 150,000 km3, GRB lavas form at least 66% of the total volume of the Columbia River Basalt Group. New 40Ar/39Ar dates for GRB lavas reveal they were emplaced within a maximum period of 0.42 ± 0.18 My. A well-documented stratigraphy indicates at least 110 GRB flow fields (or individual eruptions), and on this basis suggests an average inter-eruption hiatus of less than 4,000 years. Isotopic age-dating cannot resolve time gaps between GRB eruptions, and it is difficult to otherwise form a picture of the durations of eruptions because of non-uniform weathering in the top of flow fields and a general paucity of sediments between GR lavas. Where sediment has formed on top of the GRB, it varies in thickness from zero to 20-30 cm of silty to fine-sandy material, with occasional diatomaceous sediment. Individual GRB eruptions varied considerably in volume but many were greater than 1000 km3 in size. Most probably eruptive events were not equally spaced in time; some eruptions may have followed short periods of volcanic repose (perhaps 102 to 103 of yrs), whilst others could have been considerably longer (many 1000 s to > 104 yrs). Recent improvements in age-dating for other continental flood basalt (CFB) lava sequences have yielded estimates of total eruptive durations of less than 1 My for high-volume pulses of lava production. The GRB appears to be a similar example, where the main pulse occupied a brief period. Even allowing for moderate to long-duration pahoehoe flow field production, the amount of time the system spends in active lava-producing mode is small – less than c. 2.6% (based on eruption durations of approximately 10,000 yrs, as compared to the duration of the entire eruptive pulse of c. 420,000 yrs). A review of available 40Ar/39Ar data for the major voluminous phases of the Columbia River Basalt Group suggests that activity of the Steens Basalt-Imnaha Basalt-GRB may have, at times, been simultaneous, with obvious implications for climatic effects. Resolving intervals between successive eruptions during CFB province construction, and durations of main eruptive pulses, remains vital to determining the environmental impact of these huge eruptions

Pathway of human AS3MT arsenic methylation

A synthetic gene encoding human As(III) S-adenosylmethionine (SAM) methyltransferase (hAS3MT) was expressed, and the purified enzyme was characterized. The synthetic enzyme is considerably more active than a cDNA-expressed enzyme using endogenous reductants thioredoxin (Trx), thioredoxin reductase (TR), NADPH, and reduced glutathione (GSH). Each of the seven cysteines (the four conserved residues, Cys32, Cys61, Cys156, and Cys206, and nonconserved, Cys72, Cys85, and Cys250) was individually changed to serine. The nonconserved cysteine derivates were still active. None of the individual C32S, C61S, C156S, and C206S derivates were able to methylate As(III). However, the C32S and C61S enzymes retained the ability to methylate MAs(III). These observations suggest that Cys156 and Cys206 play a different role in catalysis than that of Cys32 and Cys61. A homology model built on the structure of a thermophilic orthologue indicates that Cys156 and Cys206 form the As(III) binding site, whereas Cys32 and Cys61 form a disulfide bond. Two observations shed light on the pathway of methylation. First, binding assays using the fluorescence of a single-tryptophan derivative indicate that As(GS)3 binds to the enzyme much faster than inorganic As(III). Second, the major product of the first round of methylation is MAs(III), not MAs(V), and remains enzyme-bound until it is methylated a second time. We propose a new pathway for hAS3MT catalysis that reconciles the hypothesis of Challenger ((1947) Sci. Prog., 35, 396-416) with the pathway proposed by Hayakawa et al. ((2005) Arch. Toxicol., 79, 183-191). The products are the more toxic and more carcinogenic trivalent methylarsenicals, but arsenic undergoes oxidation and reduction as enzyme-bound intermediates

Superconducting On-chip Fourier Transform Spectrometer

The kinetic inductance effect is strongly nonlinear with applied current in NbTiN, TiN and NbN thin films. This can be utilized to realize novel devices. We present results from transmission lines made with these materials, where DC (current) control is used to modulate the phase velocity thereby enabling on-chip spectrometers. Utility of such compact spectrometers is discussed, along with their natural connection with parametric amplifiers

Self-pulsation at 480 GHz from a two-color discrete mode laser diode

A discrete mode Fabry-Pérot laser is designed and fabricated to achieve two-color lasing. We demonstrate beating between the two laser modes and self-pulsation at 480 GHz

The Infrared Database of Extragalactic Observables from Spitzer I: the redshift catalog

This is the first of a series of papers on the Infrared Database of Extragalactic Observables from Spitzer (IDEOS). In this work we describe the identification of optical counterparts of the infrared sources detected in Spitzer Infrared Spectrograph (IRS) observations, and the acquisition and validation of redshifts. The IDEOS sample includes all the spectra from the Cornell Atlas of Spitzer/IRS Sources (CASSIS) of galaxies beyond the Local Group. Optical counterparts were identified from correlation of the extraction coordinates with the NASA Extragalactic Database (NED). To confirm the optical association and validate NED redshifts, we measure redshifts with unprecedented accuracy on the IRS spectra ({\sigma}(dz/(1+z))=0.0011) by using an improved version of the maximum combined pseudo-likelihood method (MCPL). We perform a multi-stage verification of redshifts that considers alternate NED redshifts, the MCPL redshift, and visual inspection of the IRS spectrum. The statistics is as follows: the IDEOS sample contains 3361 galaxies at redshift 0<z<6.42 (mean: 0.48, median: 0.14). We confirm the default NED redshift for 2429 sources and identify 124 with incorrect NED redshifts. We obtain IRS-based redshifts for 568 IDEOS sources without optical spectroscopic redshifts, including 228 with no previous redshift measurements. We provide the entire IDEOS redshift catalog in machine-readable formats. The catalog condenses our compilation and verification effort, and includes our final evaluation on the most likely redshift for each source, its origin, and reliability estimates.Comment: 11 pages, 6 figures, 1 table. Accepted for publication in MNRAS. Full redshift table in machine-readable format available at http://ideos.astro.cornell.edu/redshifts.htm

Exploring the anthelmintic properties of Australian native shrubs with respect to their potential role in livestock grazing systems

We measured in vitro anthelmintic activity in extracts from 85 species of Australian native shrub, with a view to identifying species able to provide a degree of worm control in grazing systems. Approximately 40% of the species showed significant activity in inhibiting development of Haemonchus contortus larvae. The most active extracts showed IC50 values of 60–300 mg/ml. Pre-incubation with polyvinylpolypyrrolidine removed the activity from some extracts, implicating tannins as the bioactive agent, while in other cases the pre-incubation had no effect, indicating the presence of other anthelmintic compounds. Plant reproductive maturity (onset of flowering or fruiting) was associated with increasing anthelmintic activity in some species. Variability was observed between plants of the same species growing in different environments, while variation between individual plants of the same species within a single field suggests the existence of distinct chemotypes. Significant activity against adult H. contortus worms in vitro was also demonstrated in a limited number of extracts tested against this life stage. Our study indicates that there is potential for Australian native shrubs to play an anthelmintic role in grazing systems, and highlights some plant biology factors which will need to be considered in order to maximize any anthelmintic effects.A. C. Kotze, J. O’Grady, J. Emms, A. F. Toovey, S. Hughes, P. Jessop, M. Bennell P. E. Vercoe and D. K. Revel