Capping protein: new insights into mechanism and regulation

Temporal and spatial control of the actin cytoskeleton are crucial for a range of eukaryotic cellular processes. Capping protein (CP), a ubiquitous highly conserved heterodimer, tightly caps the barbed (fast-growing) end of the actin filament and is an important component in the assembly of various actin structures, including the dynamic branched filament network at the leading edge of motile cells. New research into the molecular mechanism of how CP interacts with the actin filament in vitro and the function of CP in vivo, including discoveries of novel interactions of CP with other proteins, has greatly enhanced our understanding of the role of CP in regulating the actin cytoskeleton.     

Applying Climate Change Risk Management Tools to Integrate Streamflow Projections and Social Vulnerability

Ecosystems - Shifts in streamflow, due to future climate and land use change, may pose risks to nearby human communities. Projecting the spatial distribution and impacts of these risks requires...     

Amyloid cannot resist identification

The capacity to polymerize into amyloid fibrils is common to many proteins. While some proteins naturally form these fibrils to serve functional roles, amyloid is usually associated with pathogenic processes in which specific proteins aberrantly aggregate within cells or tissues. Though the contribution of amyloid fibrils to actual disease pathogenesis is not always clear, one possibility is that the titration of essential proteins from solution into aggregates contributes to the cellular degeneration common to many amyloid diseases. Using mammalian and yeast model systems, we recently showed that the common biophysical properties of amyloid aggregates—including strong resistance to dissolution—enable stringent purification and identification of both amyloid-forming and amyloid-associated proteins directly from cells. Strikingly, many proteins that were previously implicated in formation or clearance of intracellular aggregates, including several stress granule components, were found to co-aggregate with amyloid formed by a polyglutamine-expanded huntingtin fragment. This direct evaluation of proteins within aggregates can help identify new amyloid-forming proteins, as well as proteins that can indirectly contribute to disease mechanisms.     

NASCA report 2: Longitudinal study of relationship of exposure to space radiation and risk of lens opacity

The NASA Study of Cataract in Astronauts (NASCA) was designed to measure the impact of exposure to space radiation on progression rates of cortical, nuclear, and posterior subcapsular cataract in U.S. astronauts who have flown in space and comparison groups of astronauts who had not flown in space, and subjects with a history of military aviation. We present our analyses of 5 years of data with an average of 3.8 exams per subject. All subjects had digital lens images with the Nidek EAS 1000 Lens Imaging System. Because of high variability and skewness of opacity measures, nonparametric methods were used to test for association between rates of opacification and space radiation exposure. First, median regression was used to collapse longitudinal data into robust estimates of progression rates (opacity severity compare to time for each eye of each subject). To quantify and test for a radiation effect, median regression with the dependent variable being the maximum of the two slopes (OD and OS) per subject was then used, adjusting for the confounding variables of age, nutritional, and sun-exposure histories. Median regression showed evidence of an association between the rate of cortical progression in the worse eye with radiation dose and age. The estimated median progression rate from space radiation being 0.25 ± 0.13% lens area/Sv/year (P = 0.062). We found no relationship between radiation exposure and progression of aggregate area of posterior subcapsular cataract or nuclear progression rates. However, longer follow-up may be needed to further understand any impact of space radiation on progression rates for posterior subcapsular cataracts and nuclear cataracts, and to characterize changes to visual acuity.     

Literary inquiry and professional development in medicine: against abstractions.

The professional development discourse currently circulating in academic medicine owes much to the work of the American Board of Internal Medicine (ABIM) and their Project Professionalism.They identify the elements of altruism, duty, excellence, honor and integrity, accountability, and respect for others as forming the basis of professional development. While an admirable effort, Project Professionalism remains primarily an abstract set of attitudes and behaviors with a number of vignettes that are of limited use to medical educators attempting to foster the development of professionalism in medical students. This paper outlines an approach to professional development grounded in medical narratives written by physicians, including memoirs, essays, and poetry, that may help students grapple with the challenges of medicine that involve those very elements put forth by ABIM. An argument is made that literature may be a superior medium for engagement with these elements because of its ability to evoke discomfort and vulnerability in ways the case report does not.     

Ecological consequences of ontogenetic shifts in predator diet: Seasonal constraint of a behaviorally mediated indirect interaction

Predators play an important role in structuring assemblages through direct and cascading indirect effects. While there has been recent interest in how the strength and direction of trophic cascades vary spatially, seasonal variability in trophic links is seldom considered. In North Carolina, recruitment-failure of bay scallops typically occurs following the spring but not the fall spawning despite the presence in each of these seasons of predatory blue crabs. One explanation for this pattern is that in the fall, seasonally abundant predators of blue crabs reduce the foraging efficiency of crabs on scallops and thus the overall magnitude of top-down effects. Quantification of bay scallop consumption by blue crabs in closed mesocosms with or without pinfish supported the hypothesis that seasonally abundant adult pinfish indirectly increase survivorship of bay scallop recruits in fall by reducing predation by blue crabs. Despite voracious consumption of bay scallops during both the day and night in mesocosms to which only small blue crabs were added, blue crabs in mesocosms with visually-foraging adult pinfish consumed bay scallops only by night. Juvenile pinfish that dominate estuarine populations in spring did not impede consumption of bay scallops by blue crabs. In mesocosms from which animals could not emigrate, the addition of neither adult nor juvenile pinfish increased the mortality of blue crabs, indicating a behaviorally mediated interaction. Blue crabs restricted by adult pinfish to nocturnal feeding did not compensate for lost feeding time by increasing their night-time consumption of bay scallops. These results strongly suggest that greater survivorship of bay scallops in fall than spring is due to adult pinfish, potential predators of small blue crabs, restricting blue crab foraging to hours of dark. In spring, when pinfish are small and incapable of consuming blue crabs, blue crabs consume bay scallops by day and by night. Such seasonal variation in the number of trophic links in a system may have important evolutionary implications. By timing reproduction to occur in fall when the pinfish-crab-scallop cascade is in operation, bay scallops maximize recruitment.     

Operationalizing resilience for adaptive coral reef management under global environmental change

Cumulative pressures from global climate and ocean change combined with multiple regional and local‐scale stressors pose fundamental challenges to coral reef managers worldwide. Understanding how cumulative stressors affect coral reef vulnerability is critical for successful reef conservation now and in the future. In this review, we present the case that strategically managing for increased ecological resilience (capacity for stress resistance and recovery) can reduce coral reef vulnerability (risk of net decline) up to a point. Specifically, we propose an operational framework for identifying effective management levers to enhance resilience and support management decisions that reduce reef vulnerability. Building on a system understanding of biological and ecological processes that drive resilience of coral reefs in different environmental and socio‐economic settings, we present an Adaptive Resilience‐Based management (ARBM) framework and suggest a set of guidelines for how and where resilience can be enhanced via management interventions. We argue that press‐type stressors (pollution, sedimentation, overfishing, ocean warming and acidification) are key threats to coral reef resilience by affecting processes underpinning resistance and recovery, while pulse‐type (acute) stressors (e.g. storms, bleaching events, crown‐of‐thorns starfish outbreaks) increase the demand for resilience. We apply the framework to a set of example problems for Caribbean and Indo‐Pacific reefs. A combined strategy of active risk reduction and resilience support is needed, informed by key management objectives, knowledge of reef ecosystem processes and consideration of environmental and social drivers. As climate change and ocean acidification erode the resilience and increase the vulnerability of coral reefs globally, successful adaptive management of coral reefs will become increasingly difficult. Given limited resources, on‐the‐ground solutions are likely to focus increasingly on actions that support resilience at finer spatial scales, and that are tightly linked to ecosystem goods and services.     

A kinetically trapped intermediate of FK506 binding protein forms in vitro: chaperone machinery dominates protein folding in vivo

We have characterised the stability, binding and enzymatic properties of three human FK506 binding proteins (FKBP-12) differing only by the length and sequence of their N-terminus. One construct has a short hexa-his tag (6H-FKBP12); the second longer fusion protein (6HL-FKBP12) contains an additional thrombin protease cleavage site; the third has the long fusion tag removed and is essentially native FKBP-12 (cFKBP12). The proteins were purified both under native conditions and also using a refolding protocol. All three natively purified proteins have, within experimental error, the same peptidyl-prolyl isomerase (PPIase) activity (k(cat)/K(m) approximately 1 x 10(6)M(-1)s(-1)), and bind a natural inhibitor, rapamycin, with the same high affinity (K(d) approximately 6 nM). However, refolding of the protein containing the longer tag in vitro results in reduced PPIase activity (the k(cat)/K(m) was reduced from 1 x 10(6)M(-1)s(-1) to 0.81 x 10(6)M(-1)s(-1)) and a 6-fold affinity loss for rapamycin. Addition of both the long and short N-terminal his-tags slows the refolding kinetics of FKBP-12. However, the shorter his-tagged fusion protein regains fully native activity (> or =95%) while the longer regains only approximately 80-85% of native activity. Equilibrium urea denaturation titrations, isothermal titration calorimetry (ITC), analytical gel-filtration, and fluorescence binding data show that this loss of activity is not due to gross misfolding events, but is rather caused by the formation of a stable but subtly misfolded protein that has reduced peptidyl-prolyl isomerase (PPIase) activity and reduced affinity for rapamycin. The difference in behaviour between the in vitro refolded and native forms is due to the dominant role of the cellular chaperone/folding machinery.     

Structure-based discovery of a family of synthetic cyclophilin inhibitors showing a cyclosporin-A phenotype in Caenorhabditis elegans

Cyclophilins, which are found in all cellular compartments and with diverse biological roles, are now drug targets for a number of diseases including HIV infection, malaria and ischaemia. We used the database-mining program LIDAEUS and in silico screening to discover the dimedone family of inhibitors which show a conserved ‘ball and socket’ binding mode with a dimethyl group in the hydrophobic binding pocket of human cyclophilin A (CypA) mimicking a key interaction of the natural inhibitor cyclosporin A (CsA). The most potent derivative binds CypA with a K_d of 11.2 ± 9.2 μM and an IC₅₀ for activity against Caenorhabditis elegans (C. elegans) of 190 μM compared to 28 μM for CsA. These dimedone analogues provide a new scaffold for the synthesis of families of peptidomimetic molecules with potential activity against HIV, malaria, and helminth parasite infections.