Electron heating at Saturn's bow shock

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94695/1/jgra21551.pd

Knockout studies reveal an important role of <i>plasmodium</i> lipoic acid protein ligase a1 for asexual blood stage parasite survival

Lipoic acid (LA) is a dithiol-containing cofactor that is essential for the function of a-keto acid dehydrogenase complexes. LA acts as a reversible acyl group acceptor and 'swinging arm' during acyl-coenzyme A formation. The cofactor is post-translationally attached to the acyl-transferase subunits of the multienzyme complexes through the action of octanoyl (lipoyl): &lt;i&gt;N&lt;/i&gt;-octanoyl (lipoyl) transferase (LipB) or lipoic acid protein ligases (LplA). Remarkably, apicomplexan parasites possess LA biosynthesis as well as scavenging pathways and the two pathways are distributed between mitochondrion and a vestigial organelle, the apicoplast. The apicoplast-specific LipB is dispensable for parasite growth due to functional redundancy of the parasite's lipoic acid/octanoic acid ligases/transferases. In this study, we show that &lt;i&gt;LplA1&lt;/i&gt; plays a pivotal role during the development of the erythrocytic stages of the malaria parasite. Gene disruptions in the human malaria parasite &lt;i&gt;P.falciparum&lt;/i&gt; consistently were unsuccessful while in the rodent malaria model parasite &lt;i&gt;P. berghei&lt;/i&gt; the &lt;i&gt;LplA1&lt;/i&gt; gene locus was targeted by knock-in and knockout constructs. However, the &lt;i&gt;LplA1&lt;/i&gt; &lt;sup&gt;(-)&lt;/sup&gt; mutant could not be cloned suggesting a critical role of LplA1 for asexual parasite growth &lt;i&gt;in vitro&lt;/i&gt; and &lt;i&gt;in vivo&lt;/i&gt;. These experimental genetics data suggest that lipoylation during expansion in red blood cells largely occurs through salvage from the host erythrocytes and subsequent ligation of LA to the target proteins of the malaria parasite

A Systematic Screen to Discover and Analyze Apicoplast Proteins Identifies a Conserved and Essential Protein Import Factor

Parasites of the phylum Apicomplexa cause diseases that impact global health and economy. These unicellular eukaryotes possess a relict plastid, the apicoplast, which is an essential organelle and a validated drug target. However, much of its biology remains poorly understood, in particular its elaborate compartmentalization: four membranes defining four different spaces. Only a small number of organellar proteins have been identified in particular few proteins are known for non-luminal apicoplast compartments. We hypothesized that enlarging the catalogue of apicoplast proteins will contribute toward identifying new organellar functions and expand the realm of targets beyond a limited set of characterized pathways. We developed a bioinformatic screen based on mRNA abundance over the cell cycle and on phyletic distribution. We experimentally assessed 57 genes, and of 30 successful epitope tagged candidates eleven novel apicoplast proteins were identified. Of those, seven appear to target to the lumen of the organelle, and four localize to peripheral compartments. To address their function we then developed a robust system for the construction of conditional mutants via a promoter replacement strategy. We confirm the feasibility of this system by establishing conditional mutants for two selected genes – a luminal and a peripheral apicoplast protein. The latter is particularly intriguing as it encodes a hypothetical protein that is conserved in and unique to Apicomplexan parasites and other related organisms that maintain a red algal endosymbiont. Our studies suggest that this peripheral plastid protein, PPP1, is likely localized to the periplastid compartment. Conditional disruption of PPP1 demonstrated that it is essential for parasite survival. Phenotypic analysis of this mutant is consistent with a role of the PPP1 protein in apicoplast biogenesis, specifically in import of nuclear-encoded proteins into the organelle

A single in vivo-selected point mutation in the active center of Toxoplasma gondii ferredoxin-NADP+ reductase leads to an inactive enzyme with greatly enhanced affinity for ferredoxin

Electron transfer between plant-type [2Fe-2S] ferredoxin (Fd) and ferredoxin-NADP+ reductase (FNR) depends on the physical interaction between both proteins. We have applied a random mutagenesis approach with subsequent in vivo selection using the yeast two-hybrid system to obtain mutants of Toxoplasma gondii FNR with higher affinity for Fd. One mutant showed a 10-fold enhanced binding using affinity chromatography on immobilized Fd. A single serine-to-arginine exchange in the active site was responsible for its increased affinity. The mutant reductase was also enzymatically inactive. Homology modeling of the mutant FNR-Fd complex predicts substantial alterations of protein-FAD interactions in the active site of the enzyme with subsequent structural changes. Collectively, for the first time a point mutation in this important class of enzymes is described which leads to greatly enhanced affinity for its protein ligand

In situ observations of high-Mach number collisionless shocks in space plasmas

International audienceShock waves are widespread in collisionless space plasmas throughout the Universe. How particles are accelerated at these shocks has been the subject of much research attention. The dominant source of the high-energy particles that pervade our Galaxy (cosmic rays) is thought to be the high-Mach number collisionless shocks that form around young supernova remnants, but it is unclear how much the lower Mach number collisionless shock waves frequently encountered by spacecraft in Solar System space plasmas can tell us about particle acceleration in the higher Mach number regime. Here we review recent studies of the shock wave that stands in the solar wind in front of the planet Saturn (Saturn's bow shock), based on Cassini spacecraft observations. This review represents a new direction of shock physics research, with the potential to bridge the gap between Solar System and astrophysical shocks. These studies have confirmed that Saturn's bow shock is one of the strongest shocks in the Solar System, and a recent discovery indicates that electron acceleration at high-Mach numbers may occur irrespective of the upstream magnetic field geometry. This is important because astrophysical shocks can often only be studied remotely via emissions associated with accelerated electrons. We discuss possible future directions of this emerging sub-field of collisionless space plasma shock physics

Electron acceleration to relativistic energies at a strong quasi-parallel shock wave

International audienceElectrons can be accelerated to ultrarelativistic energies at strong (high-Mach number) collisionless shock waves that form when stellar debris rapidly expands after a supernova. Collisionless shock waves also form in the flow of particles from the Sun (the solar wind), and extensive spacecraft observations have established that electron acceleration at these shocks is effectively absent whenever the upstream magnetic field is roughly parallel to the shock surface normal (quasi-parallel conditions). However, it is unclear whether this magnetic dependence of electron acceleration also applies to the far stronger shocks around young supernova remnants, where local magnetic conditions are poorly understood. Here we present Cassini spacecraft observations of an unusually strong solar system shock wave (Saturn's bow shock) where significant local electron acceleration has been confirmed under quasi-parallel magnetic conditions for the first time, contradicting the established magnetic dependence of electron acceleration at solar system shocks. Furthermore, the acceleration led to electrons at relativistic energies (~MeV), comparable to the highest energies ever attributed to shock-acceleration in the solar wind. These observations suggest that at high-Mach numbers, like those of young supernova remnant shocks, quasi-parallel shocks become considerably more effective electron accelerators

The Mycobacterium tuberculosis LipB enzyme functions as a cysteine / lysine dyad acyltransferase.

Lipoic acid is essential for the activation of a number of protein complexes involved in key metabolic processes. Growth of Mycobacterium tuberculosis relies on a pathway in which the lipoate attachment group is synthesized from an endogenously produced octanoic acid moiety. In patients with multiple-drug-resistant M. tuberculosis, expression of one gene from this pathway, lipB, encoding for octanoyl-[acyl carrier protein]-protein acyltransferase is considerably up-regulated, thus making it a potential target in the search for novel antiinfectives against tuberculosis. Here we present the crystal structure of the M. tuberculosis LipB protein at atomic resolution, showing an unexpected thioether-linked active-site complex with decanoic acid. We provide evidence that the transferase functions as a cysteine/lysine dyad acyltransferase, in which two invariant residues (Lys-142 and Cys-176) are likely to function as acid/base catalysts. Analysis by MS reveals that the LipB catalytic reaction proceeds by means of an internal thioesteracyl intermediate. Structural comparison of LipB with lipoate protein ligase A indicates that, despite conserved structural and sequence active-site features in the two enzymes, 4'-phosphopantetheine-bound octanoic acid recognition is a specific property of LipB

Outer magnetospheric structure: Jupiter and Saturn compared

The Jovian dayside magnetosphere is traditionally divided into three different regions with the outermost region, colloquially referred to as the cushion region, existing between the outer edge of the magnetodisk and the magnetopause. Magnetometer and plasma data from 6 different spacecraft are used to determine the average properties of this region, including its characteristic thickness at the subsolar point, and these observations are compared with data from the Saturnian magnetosphere obtained using the Pioneer, Voyager, and Cassini spacecraft. Significant differences are found in the structure of the two rotationally driven magnetospheres with the Saturnian system showing little evidence for the cushion region seen at Jupiter. These differences are discussed in terms of the parameter regimes pertinent to each planet, and the potential effect of magnetodisk warping at Saturn is discussed. It is tentatively suggested that while the Jovian magnetodisk typically breaks down several tens of planetary radii inside the magnetopause, thus allowing plasma-depleted flux tubes beyond it to relax into the cushion region configuration, the Saturnian magnetodisk may persist until much closer to the magnetospheric boundary. A number of observational tests of this hypothesis are proposed, and the need for improved observations at both planets is stressed