Alpha-2-Macroglobulin, a Hypochlorite-Regulated Chaperone and Immune System Modulator

Alpha-macroglobulins are ancient proteins that include monomeric, dimeric, and tetrameric family members. In humans, and many other mammals, the predominant alpha-macroglobulin is alpha-2-macroglobulin (α2M), a tetrameric protein that is constitutively abundant in biological fluids (e.g., blood plasma, cerebral spinal fluid, synovial fluid, ocular fluid, and interstitial fluid). α2M is best known for its remarkable ability to inhibit a broad spectrum of proteases, but the full gamut of its activities affects diverse biological processes. For example, α2M can stabilise and facilitate the clearance of the Alzheimer’s disease-associated amyloid beta (Aβ) peptide. Additionally, α2M can influence the signalling of cytokines and growth factors including neurotrophins. The results of several studies support the idea that the functions of α2M are uniquely regulated by hypochlorite, an oxidant that is generated during inflammation, which induces the native α2M tetramer to dissociate into dimers. This review will discuss the evidence for hypochlorite-induced regulation of α2M and the possible implications of this in neuroinflammation and neurodegeneration

Spatially Resolved Images of Dust Belt(s) Around the Planet-hosting Subgiant Kappa CrB

We present Herschel spatially resolved images of the debris disc orbiting the subgiant Kappa CrB. Not only are these the first resolved images of a debris disc orbiting a subgiant, but Kappa CrB is a rare example of an intermediate mass star where a detailed study of the structure of the planetary system can be made, including both planets and planetesimal belt(s). The only way to discover planets around such stars using the radial velocity technique is to observe 'retired' A stars, which are cooler and slower rotators compared to their main-sequence counterparts. A planetary companion has already been detected orbiting the subgiant Kappa CrB, with revised parameters of m sin i = 2.1MJ and apl = 2.8AU (Johnson et al. 2008a). We present additional Keck I HIRES radial velocity measurements that provide evidence for a second planetary companion, alongside Keck II AO imaging that places an upper limit on the mass of this companion. Modelling of our Herschel images shows that the dust is broadly distributed, but cannot distinguish between a single wide belt (from 20 to 220AU) or two narrow dust belts (at around 40 and 165AU). Given the existence of a second planetary companion beyond approximately 3AU it is possible that the absence of dust within approximately 20AU is caused by dynamical depletion, although the observations are not inconsistent with depletion of these regions by collisional erosion, which occurs at higher rates closer to the star.Comment: Updated abstrac

Secondary gas in debris discs released following the decay of long-lived radioactive nuclides, catastrophic or resurfacing collisions

Kuiper-like belts of planetesimals orbiting stars other than the Sun are most commonly detected from the thermal emission of small dust produced in collisions. Emission from gas, most notably CO, highlights the cometary nature of these planetesimals. Here we present models for the release of gas from comet-like bodies in these belts, both due to their thermophysical evolution, most notably the decay of long-lived radioactive nuclides and collisional evolution, including catastrophic and gentler resurfacing collisions. We show that the rate of gas release is not proportional to the rate of dust release, if non-catastrophic collisions or thermal evolution dominate the release of CO gas. In this case, care must be taken when inferring the composition of comets. Non-catastrophic collisions dominate the gas production at earlier times than catastrophic collisions, depending on the properties of the planetesimal belt. We highlight the importance of the thermal evolution of comets, including crucially the decay of long-lived radioactive nuclides, as a source of CO gas around young (<50Myr) planetary systems, if large (10-100s kms) planetesimals are present.Comment: Submitted to MNRAS, 16 page

Protease-activated alpha-2-macroglobulin can inhibit amyloid formation via two distinct mechanisms.

α(2)-Macroglobulin (α(2)M) is an extracellular chaperone that inhibits amorphous and fibrillar protein aggregation. The reaction of α(2)M with proteases results in an 'activated' conformation, where the proteases become covalently-linked within the interior of a cage-like structure formed by α(2)M. This study investigates, the effect of activation on the ability of α(2)M to inhibit amyloid formation by Aβ(1-42) and I59T human lysozyme and shows that protease-activated α(2)M can act via two distinct mechanisms: (i) by trapping proteases that remain able to degrade polypeptide chains and (ii) by a chaperone action that prevents misfolded clients from continuing along the amyloid forming pathway

Spatially resolved images of dust belt(s) around the planet-hosting subgiant κ CrB

We present Herschel spatially resolved images of the debris disc orbiting the subgiant κ Coronae Borealis (κ CrB). Not only are these the first resolved images of a debris disc orbiting a subgiant, but κ CrB is a rare example of an intermediate mass star where a detailed study of the structure of the planetary system can be made, including both planets and planetesimal belt(s). The only way to discover planets using the radial velocity technique around such stars is to observe ‘retired’ A stars, which are cooler and slower rotators compared to their main-sequence counterparts. A planetary companion has already been detected orbiting the subgiant κ CrB, with revised parameters of msin i = 2.1 M_J and a_(pl) = 2.8 au (Johnson et al. 2008). We present additional Keck I HIRES (High Resolution Echelle Spectrometer) radial velocity measurements that provide evidence for a second planetary companion, alongside Keck II adaptive optics imaging that places an upper limit on the mass of this companion. Modelling of our Herschel images shows that the dust is broadly distributed, but cannot distinguish between a single wide belt (from 20 to 220 au) or two narrow dust belts (at around 40 and 165 au). Given the existence of a second planetary companion beyond ∼3 au it is possible that the absence of dust within ∼20 au is caused by dynamical depletion, although the observations are not inconsistent with depletion of these regions by collisional erosion, which occurs at higher rates closer to the star

Resistant starch and exercise independently attenuate weight regain on a high fat diet in a rat model of obesity

<p>Abstract</p> <p>Background</p> <p>Long-term weight reduction remains elusive for many obese individuals. Resistant starch (RS) and exercise may be useful for weight maintenance. The effects of RS, with or without exercise, on weight regain was examined during relapse to obesity on a high carbohydrate, high fat (HC/HF) diet.</p> <p>Methods</p> <p>Obesity-prone rats were fed <it>ad libitum </it>for 16 weeks then weight reduced on a low fat diet to induce a 17% body weight loss (weight reduced rats). Weight reduced rats were maintained on an energy-restricted low fat diet for 18 weeks, with or without a daily bout of treadmill exercise. Rats were then allowed free access to HC/HF diet containing low (0.3%) or high (5.9%) levels of RS. Weight regain, energy balance, body composition, adipocyte cellularity, and fuel utilization were monitored as rats relapsed to obesity and surpassed their original, obese weight.</p> <p>Results</p> <p>Both RS and exercise independently attenuated weight regain by reducing the energy gap between the drive to eat and suppressed energy requirements. Exercise attenuated the deposition of lean mass during relapse, whereas its combination with RS sustained lean mass accrual as body weight returned. Early in relapse, RS lowered insulin levels and reduced the deposition of fat in subcutaneous adipose tissue. Exercise cessation at five weeks of relapse led to increased weight gain, body fat, subcutaneous adipocytes, and decreased lean mass; all detrimental consequences to overall metabolic health.</p> <p>Conclusions</p> <p>These data are the first to show the complimentary effects of dietary RS and regular exercise in countering the metabolic drive to regain weight following weight loss and suggest that exercise cessation, in the context of relapse on a HC/HF diet, may have dire metabolic consequences.</p

High-Throughput Analysis of Lung Immune Cells in a Combined Murine Model of Agriculture Dust-Triggered Airway Inflammation With Rheumatoid Arthritis

Rheumatoid arthritis (RA)-associated lung disease is a leading cause of mortality in RA, yet the mechanisms linking lung disease and RA remain unknown. Using an established murine model of RA-associated lung disease combining collagen-induced arthritis (CIA) with organic dust extract (ODE)-induced airway inflammation, differences among lung immune cell populations were analyzed by single cell RNA-sequencing. Additionally, four lung myeloid-derived immune cell populations including macrophages, monocytes/macrophages, monocytes, and neutrophils were isolated by fluorescence cell sorting and gene expression was determined by NanoString analysis. Unsupervised clustering revealed 14 discrete clusters among Sham, CIA, ODE, and CIA+ODE treatment groups: 3 neutrophils (inflammatory, resident/transitional, autoreactive/suppressor), 5 macrophages (airspace, differentiating/recruited, recruited, resident/interstitial, and proliferative airspace), 2 T-cells (differentiating and effector), and a single cluster each of inflammatory monocytes, dendritic cells, B-cells and natural killer cells. Inflammatory monocytes, autoreactive/suppressor neutrophils, and recruited/differentiating macrophages were predominant with arthritis induction (CIA and CIA+ODE). By specific lung cell isolation, several interferon-related and autoimmune genes were disproportionately expressed among CIA and CIA+ODE (e.g. Oasl1, Oas2, Ifit3, Gbp2, Ifi44, and Zbp1), corresponding to RA and RA-associated lung disease. Monocytic myeloid-derived suppressor cells were reduced, while complement genes (e.g. C1s1 and Cfb) were uniquely increased in CIA+ODE mice across cell populations. Recruited and inflammatory macrophages/monocytes and neutrophils expressing interferon-, autoimmune-, and complement-related genes might contribute towards pro-fibrotic inflammatory lung responses following airborne biohazard exposures in setting of autoimmune arthritis and could be predictive and/or targeted to reduce disease burden

Human pregnancy zone protein stabilizes misfolded proteins including preeclampsia- and Alzheimer's-associated amyloid beta peptide.

Protein misfolding underlies the pathology of a large number of human disorders, many of which are age-related. An exception to this is preeclampsia, a leading cause of pregnancy-associated morbidity and mortality in which misfolded proteins accumulate in body fluids and the placenta. We demonstrate that pregnancy zone protein (PZP), which is dramatically elevated in maternal plasma during pregnancy, efficiently inhibits in vitro the aggregation of misfolded proteins, including the amyloid beta peptide (Aβ) that is implicated in preeclampsia as well as with Alzheimer's disease. The mechanism by which this inhibition occurs involves the formation of stable complexes between PZP and monomeric Aβ or small soluble Aβ oligomers formed early in the aggregation pathway. The chaperone activity of PZP is more efficient than that of the closely related protein alpha-2-macroglobulin (α2M), although the chaperone activity of α2M is enhanced by inducing its dissociation into PZP-like dimers. By immunohistochemistry analysis, PZP is found primarily in extravillous trophoblasts in the placenta. In severe preeclampsia, PZP-positive extravillous trophoblasts are adjacent to extracellular plaques containing Aβ, but PZP is not abundant within extracellular plaques. Our data support the conclusion that the up-regulation of PZP during pregnancy represents a major maternal adaptation that helps to maintain extracellular proteostasis during gestation in humans. We propose that overwhelming or disrupting the chaperone function of PZP could underlie the accumulation of misfolded proteins in vivo. Attempts to characterize extracellular proteostasis in pregnancy will potentially have broad-reaching significance for understanding disease-related protein misfolding.Wellcome Trust Programme Grant 094425/Z/10/

Alpha-2-Macroglobulin Is Acutely Sensitive to Freezing and Lyophilization: Implications for Structural and Functional Studies.

Alpha-2-macroglobulin is an abundant secreted protein that is of particular interest because of its diverse ligand binding profile and multifunctional nature, which includes roles as a protease inhibitor and as a molecular chaperone. The activities of alpha-2-macroglobulin are typically dependent on whether its conformation is native or transformed (i.e. adopts a more compact conformation after interactions with proteases or small nucleophiles), and are also influenced by dissociation of the native alpha-2-macroglobulin tetramer into stable dimers. Alpha-2-macroglobulin is predominately present as the native tetramer in vivo; once purified from human blood plasma, however, alpha-2-macroglobulin can undergo a number of conformational changes during storage, including transformation, aggregation or dissociation. We demonstrate that, particularly in the presence of sodium chloride or amine containing compounds, freezing and/or lyophilization of alpha-2-macroglobulin induces conformational changes with functional consequences. These conformational changes in alpha-2-macroglobulin are not always detected by standard native polyacrylamide gel electrophoresis, but can be measured using bisANS fluorescence assays. Increased surface hydrophobicity of alpha-2-macroglobulin, as assessed by bisANS fluorescence measurements, is accompanied by (i) reduced trypsin binding activity, (ii) increased chaperone activity, and (iii) increased binding to the surfaces of SH-SY5Y neurons, in part, via lipoprotein receptors. We show that sucrose (but not glycine) effectively protects native alpha-2-macroglobulin from denaturation during freezing and/or lyophilization, thereby providing a reproducible method for the handling and long-term storage of this protein.Early Career Fellowship from the National Health and Medical Research Council GNT1012521(A.R.W.); Wellcome Trust Programme Grant (J.R.K., C.M.D.) 094425/Z/10/Z; Samsung GRO Grant (M.R.W.)This is the final version of the article. It first appeared from PLoS via http://dx.doi.org/10.1371/journal.pone.013003