A revised biosynthetic pathway for the cofactor F-420 in prokaryotes

Cofactor F420 plays critical roles in primary and secondary metabolism in a range of bacteria and archaea as a low-potential hydride transfer agent. It mediates a variety of important redox transformations involved in bacterial persistence, antibiotic biosynthesis, pro-drug activation and methanogenesis. However, the biosynthetic pathway for F420 has not been fully elucidated: neither the enzyme that generates the putative intermediate 2-phospho-L-lactate, nor the function of the FMN-binding C-terminal domain of the γ-glutamyl ligase (FbiB) in bacteria are known. Here we present the structure of the guanylyltransferase FbiD and show that, along with its archaeal homolog CofC, it accepts phosphoenolpyruvate, rather than 2-phospho-L-lactate, as the substrate, leading to the formation of the previously uncharacterized intermediate dehydro-F420-0. The C-terminal domain of FbiB then utilizes FMNH2 to reduce dehydro-F420-0, which produces mature F420 species when combined with the γ-glutamyl ligase activity of the N-terminal domain. These new insights have allowed the heterologous production of F420 from a recombinant F420 biosynthetic pathway in Escherichia coli

The Crab pulsar and its red knot in the near-infrared

We present near-infrared observations obtained with ISAAC on the VLT of the Crab pulsar and its environment. Photometry of the pulsar in Js, H and Ks shows the pulsar spectrum to extend fairly smoothly from the UV/optical regime. PSF subtraction of the pulsar allows us to study its immediate neighborhood in some detail. In particular, the knot positioned just 0.6 arcsec from the pulsar has been revealed in the IR. Using also archival HST data for the knot, we have measured its broad band spectrum to rise steeply into the IR, in contrast to the spectrum of the pulsar itself.Comment: Accepted by A&

The relative and absolute timing accuracy of the EPIC-pn camera on XMM-Newton, from X-ray pulsations of the Crab and other pulsars

Reliable timing calibration is essential for the accurate comparison of XMM-Newton light curves with those from other observatories, to ultimately use them to derive precise physical quantities. The XMM-Newton timing calibration is based on pulsar analysis. However, as pulsars show both timing noise and glitches, it is essential to monitor these calibration sources regularly. To this end, the XMM-Newton observatory performs observations twice a year of the Crab pulsar to monitor the absolute timing accuracy of the EPIC-pn camera in the fast Timing and Burst modes. We present the results of this monitoring campaign, comparing XMM-Newton data from the Crab pulsar (PSR B0531+21) with radio measurements. In addition, we use five pulsars (PSR J0537-69, PSR B0540-69, PSR B0833-45, PSR B1509-58 and PSR B1055-52) with periods ranging from 16 ms to 197 ms to verify the relative timing accuracy. We analysed 38 XMM-Newton observations (0.2-12.0 keV) of the Crab taken over the first ten years of the mission and 13 observations from the five complementary pulsars. All the data were processed with the SAS, the XMM-Newton Scientific Analysis Software, version 9.0. Epoch folding techniques coupled with \chi^{2} tests were used to derive relative timing accuracies. The absolute timing accuracy was determined using the Crab data and comparing the time shift between the main X-ray and radio peaks in the phase folded light curves. The relative timing accuracy of XMM-Newton is found to be better than 10^{-8}. The strongest X-ray pulse peak precedes the corresponding radio peak by 306\pm9 \mus, which is in agreement with other high energy observatories such as Chandra, INTEGRAL and RXTE. The derived absolute timing accuracy from our analysis is \pm48 \mus.Comment: 16 pages, 9 figures. Accepted for publication on A&

Desacetyl-α-melanocyte stimulating hormone and α-melanocyte stimulating hormone are required to regulate energy balance.

OBJECTIVE: Regulation of energy balance depends on pro-opiomelanocortin (POMC)-derived peptides and melanocortin-4 receptor (MC4R). Alpha-melanocyte stimulating hormone (α-MSH) is the predicted natural POMC-derived peptide that regulates energy balance. Desacetyl-α-MSH, the precursor for α-MSH, is present in brain and blood. Desacetyl-α-MSH is considered to be unimportant for regulating energy balance despite being more potent (compared with α-MSH) at activating the appetite-regulating MC4R in vitro. Thus, the physiological role for desacetyl-α-MSH is still unclear. METHODS: We created a novel mouse model to determine whether desacetyl-α-MSH plays a role in regulating energy balance. We engineered a knock in targeted QKQR mutation in the POMC protein cleavage site that blocks the production of both desacetyl-α-MSH and α-MSH from adrenocorticotropin (ACTH1-39). RESULTS: The mutant ACTH1-39 (ACTHQKQR) functions similar to native ACTH1-39 (ACTHKKRR) at the melanocortin 2 receptor (MC2R) in vivo and MC4R in vitro. Male and female homozygous mutant ACTH1-39 (Pomctm1/tm1) mice develop the characteristic melanocortin obesity phenotype. Replacement of either desacetyl-α-MSH or α-MSH over 14 days into Pomctm1/tm1 mouse brain significantly reverses excess body weight and fat mass gained compared to wild type (WT) (Pomcwt/wt) mice. Here, we identify both desacetyl-α-MSH and α-MSH peptides as regulators of energy balance and highlight a previously unappreciated physiological role for desacetyl-α-MSH. CONCLUSIONS: Based on these data we propose that there is potential to exploit the naturally occurring POMC-derived peptides to treat obesity but this relies on first understanding the specific function(s) for desacetyl-α-MSH and α-MSH

Optical photometry of the PSR B0656+14 and its neighborhood

We present the results of the broad-band photometry of the nearby middle-aged radio pulsar PSR B0656+14 and its neighborhood obtained with the 6-meter telescope of the SAO RAS and with the Hubble Space Telescope. The broad-band spectral flux $F_\nu$ of the pulsar decreases with increasing frequency in the near-IR range and increases with frequency in the near-UV range. The increase towards UV can be naturally interpreted as the Rayleigh-Jeans tail of the soft thermal component of the X-ray spectrum emitted from the surface of the cooling neutron star. Continuation of the power-law component, which dominates in the high-energy tail of the X-ray spectrum, to the IR-optical-UV frequencies is consistent with the observed fluxes. This suggests that the non-thermal pulsar radiation may be of the same origin in a broad frequency range from IR to hard X-rays. We also studied 4 objects detected in the pulsar's 5" neighborhood.Comment: 12 pages, 20 figures, submitted to A&A. Images are available in FITS format at http://www.ioffe.rssi.ru/astro/NSG/obs/0656-phot.htm

Svestka's Research: Then and Now

Zdenek Svestka's research work influenced many fields of solar physics, especially in the area of flare research. In this article I take five of the areas that particularly interested him and assess them in a "then and now" style. His insights in each case were quite sound, although of course in the modern era we have learned things that he could not readily have envisioned. His own views about his research life have been published recently in this journal, to which he contributed so much, and his memoir contains much additional scientific and personal information (Svestka, 2010).Comment: Invited review for "Solar and Stellar Flares," a conference in honour of Prof. Zden\v{e}k \v{S}vestka, Prague, June 23-27, 2014. This is a contribution to a Topical Issue in Solar Physics, based on the presentations at this meeting (Editors Lyndsay Fletcher and Petr Heinzel

A slow-forming isopeptide bond in the structure of the major pilin SpaD from Corynebacterium diphtheriae has implications for pilus assembly

The Gram-positive organism Corynebacterium diphtheriae, the cause of diphtheria in humans, expresses pili on its surface which it uses for adhesion and colonization of its host. These pili are covalent protein polymers composed of three types of pilin subunit that are assembled by specific sortase enzymes. A structural analysis of the major pilin SpaD, which forms the polymeric backbone of one of the three types of pilus expressed by C. diphtheriae, is reported. Mass-spectral and crystallographic analysis shows that SpaD contains three internal Lys-Asn isopeptide bonds. One of these, shown by mass spectrometry to be located in the N-terminal D1 domain of the protein, only forms slowly, implying an energy barrier to bond formation. Two crystal structures, of the full-length three-domain protein at 2.5Å resolution and of a two-domain (D2-D3) construct at 1.87Å resolution, show that each of the three Ig-like domains contains a single Lys-Asn isopeptide-bond cross-link, assumed to give mechanical stability as in other such pili. Additional stabilizing features include a disulfide bond in the D3 domain and a calcium-binding loop in D2. The N-terminal D1 domain is more flexible than the others and, by analogy with other major pilins of this type, the slow formation of its isopeptide bond can be attributed to its location adjacent to the lysine used in sortase-mediated polymerization during pilus assembly.open0

Evaluating the boundary and covering degree of planar Minkowski sums and other geometrical convolutions

AbstractAlgorithms are developed, based on topological principles, to evaluate the boundary and “internal structure” of the Minkowski sum of two planar curves. A graph isotopic to the envelope curve is constructed by computing its characteristic points. The edges of this graph are in one-to-one correspondence with a set of monotone envelope segments. A simple formula allows a degree to be assigned to each face defined by the graph, indicating the number of times its points are covered by the Minkowski sum. The boundary can then be identified with the set of edges that separate faces of zero and non-zero degree, and the boundary segments corresponding to these edges can be approximated to any desired geometrical accuracy. For applications that require only the Minkowski sum boundary, the algorithm minimizes geometrical computations on the “internal” envelope edges, that do not contribute to the final boundary. In other applications, this internal structure is of interest, and the algorithm provides comprehensive information on the covering degree for different regions within the Minkowski sum. Extensions of the algorithm to the computation of Minkowski sums in R3, and other forms of geometrical convolution, are briefly discussed

High-time Resolution Astrophysics and Pulsars

The discovery of pulsars in 1968 heralded an era where the temporal characteristics of detectors had to be reassessed. Up to this point detector integration times would normally be measured in minutes rather seconds and definitely not on sub-second time scales. At the start of the 21st century pulsar observations are still pushing the limits of detector telescope capabilities. Flux variations on times scales less than 1 nsec have been observed during giant radio pulses. Pulsar studies over the next 10 to 20 years will require instruments with time resolutions down to microseconds and below, high-quantum quantum efficiency, reasonable energy resolution and sensitive to circular and linear polarisation of stochastic signals. This chapter is review of temporally resolved optical observations of pulsars. It concludes with estimates of the observability of pulsars with both existing telescopes and into the ELT era.Comment: Review; 21 pages, 5 figures, 86 references. Book chapter to appear in: D.Phelan, O.Ryan & A.Shearer, eds.: High Time Resolution Astrophysics (Astrophysics and Space Science Library, Springer, 2007). The original publication will be available at http://www.springerlink.co