The structure of human 15-lipoxygenase-2 with a substrate mimic

Atherosclerosis is associated with chronic inflammation occurring over decades. The enzyme 15-lipoxygenase-2 (15-LOX-2) is highly expressed in large atherosclerotic plaques, and its activity has been linked to the progression of macrophages to the lipid-laden foam cells present in atherosclerotic plaques.We report here the crystal structure of human 15-LOX-2 in complex with an inhibitor that appears to bind as a substrate mimic. 15-LOX-2 contains a long loop, composed of hydrophobic amino acids, which projects from the amino-terminal membrane-binding domain. The loop is flanked by two Ca2+-binding sites that confer Ca2+-dependent membrane binding. A comparison of the human 15-LOX-2 and 5-LOX structures reveals similarities at the active sites, as well striking differences that can be exploited for design of isoform-selective inhibitors. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc

Monte Carlo investigation of the magnetic anisotropy in Fe/Dy multilayers

By Monte Carlo simulations in the canonical ensemble, we have studied the magnetic anisotropy in Fe/Dy amorphous multilayers. This work has been motivated by experimental results which show a clear correlation between the magnetic perpendicular anisotropy and the substrate temperature during elaboration of the samples. Our aim is to relate macroscopic magnetic properties of the multilayers to their structure, more precisely their concentration profile. Our model is based on concentration dependent exchange interactions and spin values, on random magnetic anisotropy and on the existence of locally ordered clusters that leads to a perpendicular magnetisation. Our results evidence that a compensation point occurs in the case of an abrupt concentration profile. Moreover, an increase of the noncollinearity of the atomic moments has been evidenced when the Dy anisotropy constant value grows. We have also shown the existence of inhomogeneous magnetisation profiles along the samples which are related to the concentration profiles

Sulfatides Partition Disabled-2 in Response to Platelet Activation

Background: Platelets contact each other at the site of vascular injury to stop bleeding. One negative regulator of platelet aggregation is Disabled-2 (Dab2), which is released to the extracellular surface upon platelet activation. Dab2 inhibits platelet aggregation through its phosphotyrosine-binding (PTB) domain by competing with fibrinogen for aIIbb3 integrin receptor binding by an unknown mechanism. Methodology/Principal Findings: Using protein-lipid overlay and liposome-binding assays, we identified that the N-terminal region of Dab2, including its PTB domain (N-PTB), specifically interacts with sulfatides. Moreover, we determined that such interaction is mediated by two conserved basic motifs with a dissociation constant (K d) of 0.6 mM as estimated by surface plasmon resonance (SPR) analysis. In addition, liposome-binding assays combined with mass spectroscopy studies revealed that thrombin, a strong platelet agonist, cleaved N-PTB at a site located between the basic motifs, a region that becomes protected from thrombin cleavage when bound to sulfatides. Sulfatides on the platelet surface interact with coagulation proteins, playing a major role in haemostasis. Our results show that sulfatides recruit N-PTB to the platelet surface, sequestering it from integrin receptor binding during platelet activation. This is a transient recruitment that follows N-PTB internalization by an actin-dependent process. Conclusions/Significance: Our experimental data support a model where two pools of Dab2 co-exist at the platelet surface

Conformation of the Solute-Binding Protein AdcAII Influences Zinc Uptake in Streptococcus pneumoniae.

Streptococcus pneumoniae scavenges essential zinc ions from the host during colonization and infection. This is achieved by the ATP-binding cassette transporter, AdcCB, and two solute-binding proteins (SBPs), AdcA and AdcAII. It has been established that AdcAII serves a greater role during initial infection, but the molecular details of how the protein selectively acquires Zn(II) remain poorly understood. This can be attributed to the refractory nature of metal-free AdcAII to high-resolution structural determination techniques. Here, we overcome this issue by separately mutating the Zn(II)-coordinating residues and performing a combination of structural and biochemical analyses on the variant proteins. Structural analyses of Zn(II)-bound AdcAII variants revealed that specific regions within the protein underwent conformational changes via direct coupling to each of the metal-binding residues. Quantitative in vitro metal-binding assays combined with affinity determination and phenotypic growth assays revealed that each of the four Zn(II)-coordinating residues contributes to metal binding by AdcAII. Intriguingly, the phenotypic growth impact of the mutant adcAII alleles was, in general, independent of affinity, suggesting that the Zn(II)-bound conformation of the SBP is crucial for efficacious metal uptake. Collectively, these data highlight the intimate coupling of ligand affinity with protein conformational change in ligand-receptor proteins and provide a putative mechanism for AdcAII. These findings provide further mechanistic insight into the structural and functional diversity of SBPs that is broadly applicable to other prokaryotes

The three-dimensional structure of the biotin carboxylase-biotin carboxyl carrier protein complex of E. coli acetyl-CoA carboxylase

Acetyl-coenzyme A (acetyl-CoA) carboxylase is a biotin-dependent, multifunctional enzyme that catalyzes the regulated step in fatty acid synthesis. The Escherichia coli enzyme is composed of a homodimeric biotin carboxylase (BC), biotinylated biotin carboxyl carrier protein (BCCP), and an α2β2 heterotetrameric carboxyltransferase. This enzyme complex catalyzes two half-reactions to form malonyl-coenzyme A. BC and BCCP participate in the first half-reaction, whereas carboxyltransferase and BCCP are involved in the second. Three-dimensional structures have been reported for the individual subunits; however, the structural basis for how BCCP reacts with the carboxylase or transferase is unknown. Therefore, we report here the crystal structure of E. coli BCCP complexed with BC to a resolution of 2.49 Å. The protein-protein complex shows a unique quaternary structure and two distinct interfaces for each BCCP monomer. These BCCP binding sites are unique compared to phylogenetically related biotin-dependent carboxylases and therefore provide novel targets for developing antibiotics against bacterial acetyl-CoA carboxylase. © 2013 Elsevier Ltd

Holocene vegetation and climate history in Baikal Siberia reconstructed from pollen records and its implications for archaeology

Past research has greatly improved our understanding of palaeoenvironmental changes in the Lake Baikal Region, but at the same time has indicated intra-regional variations in this vast study area. Here we present a new AMS-dated late glacial-middle Holocene (ca. 13,500-4000 cal. yr BP) pollen record from Lake Ochaul (54 degrees 14'N, 106 degrees 28'E; altitude 641 m a.s.l.) situated in the less-studied area of Cis-Baikal and compare reconstructed vegetation and climate dynamics with the published environmental history of Trans-Baikal based on the pollen record from Lake Kotokel (52 degrees 47'N, 108 degrees 07'E; altitude 458 m a.s.l.). Although both records show comparable major long-term trends in vegetation, there are considerable differences. Around Ochaul the landscape was relatively open during the Younger Dryas stadial, but forest vegetation started to spread at the late glacial/Holocene transition (ca. 11,650 cal. yr BP), thus ca. 1000 years earlier than around Kotokel. While in both regions taiga forests spread during the early and middle Holocene, the marked increase in Scots pine pollen in the Kotokel record after ca. 6800 cal. yr BP is not seen in that from Ochaul, where birch and coniferous taxa, such as Siberian pine, larch, spruce and fir, dominate, indicating different environmental conditions and driving forces in both study regions. However, the pollen data from Ochaul emphasizes that the Cis-Baikal area also saw a continuous increase in forest cover and in the proportion of conifers over birch trees and shrubs during the early-middle Holocene, which may have contributed to a decrease in the number of large herbivores, the main food resource of the Early Neolithic hunter-gatherer groups. This and rather abrupt reorganization of atmospheric circulation, which affected atmospheric precipitation distribution resulting in thicker and longer-lasting snow cover, may have led to a collapse of Early Neolithic Kitoi populations ca. 6660 cal. yr BP followed by a cultural "hiatus" in the archaeological records during the Middle Neolithic phase (ca. 6660-6060 cal. yr BP). The results stress the importance of sub-regional palaeoenvironmental studies and the need for a representative network of well-dated, high-resolution sediment archives for a better understanding of environmental changes and their potential impacts on the hunter-gatherer populations in the archaeologically-defined micro-regions

Shopping centre siting and modal choice in Belgium: a destination based analysis

Although modal split is only one of the elements considered in decision-making on new shopping malls, it remarkably often arises in arguments of both proponents and opponents. Today, this is also the case in the debate on the planned development of three major shopping malls in Belgium. Inspired by such debates, the present study focuses on the impact of the location of shopping centres on the travel mode choice of the customers. Our hypothesis is that destination-based variables such as embeddedness in the urban fabric, accessibility and mall size influence the travel mode choice of the visitors. Based on modal split data and location characteristics of seventeen existing shopping centres in Belgium, we develop a model for a more sustainable siting policy. The results show a major influence of the location of the shopping centre in relation to the urban form, and of the size of the mall. Shopping centres that are part of a dense urban fabric, measured through population density, are less car dependent. Smaller sites will attract more cyclists and pedestrians. Interestingly, our results deviate significantly from the figures that have been put forward in public debates on the shopping mall issue in Belgium