The Acidic Domains of the Toc159 Chloroplast Preprotein Receptor Family are Instrinsically Disordered Protein Domains

Background: The Toc159 family of proteins serve as receptors for chloroplast-destined preproteins. They directly bind to transit peptides, and exhibit preprotein substrate selectivity conferred by an unknown mechanism. The Toc159 receptors each include three domains: C-terminal membrane, central GTPase, and N-terminal acidic (A-) domains. Although the function(s) of the A-domain remains largely unknown, the amino acid sequences are most variable within these domains, suggesting they may contribute to the functional specificity of the receptors. Results: The physicochemical properties of the A-domains are characteristic of intrinsically disordered proteins (IDPs). Using CD spectroscopy we show that the A-domains of two Arabidopsis Toc159 family members (atToc132 and atToc159) are disordered at physiological pH and temperature and undergo conformational changes at temperature and pH extremes that are characteristic of IDPs. Conclusions: Identification of the A-domains as IDPs will be important for determining their precise function(s), and suggests a role in protein-protein interactions, which may explain how these proteins serve as receptors for such a wide variety of preprotein substrates

Dust devils on Earth and Mars: Extension of particle threshold laboratory simulations

Abstract not available

The acidic domains of the Toc159 chloroplast preprotein receptor family are intrinsically disordered protein domains

<p>Abstract</p> <p>Background</p> <p>The Toc159 family of proteins serve as receptors for chloroplast-destined preproteins. They directly bind to transit peptides, and exhibit preprotein substrate selectivity conferred by an unknown mechanism. The Toc159 receptors each include three domains: C-terminal membrane, central GTPase, and N-terminal acidic (A-) domains. Although the function(s) of the A-domain remains largely unknown, the amino acid sequences are most variable within these domains, suggesting they may contribute to the functional specificity of the receptors.</p> <p>Results</p> <p>The physicochemical properties of the A-domains are characteristic of intrinsically disordered proteins (IDPs). Using CD spectroscopy we show that the A-domains of two <it>Arabidopsis </it>Toc159 family members (atToc132 and atToc159) are disordered at physiological pH and temperature and undergo conformational changes at temperature and pH extremes that are characteristic of IDPs.</p> <p>Conclusions</p> <p>Identification of the A-domains as IDPs will be important for determining their precise function(s), and suggests a role in protein-protein interactions, which may explain how these proteins serve as receptors for such a wide variety of preprotein substrates.</p

Trance Logic, Age Regression, and Incomplete Responding: A Preliminary Investigation of Contextual Influences

Fifty-four hypnotizable and 49 unhypnotizable simulating subjects were age regressed to age five and asked to write the sentence “I am participating in a psychological experiment” embedded in either “standard” suggestions for regression or in the context of a suggested dream. As predicted, hypnotizable subjects were sensitive to the context manipulation (i.e., dream vs. standard), evidencing more correct or “adult” spelling (“trance logic”) during age regression when the dream context encouraged a melding of fantasy and reality. Simulators’ performance was stable across conditions. Consistent with the hypothesis that incomplete responding is at the heart of incongruous spelling during age regression, the hypnotizable and simulating subjects who evinced the least compelling experiences of age regression were the most likely to spell “adult” words correctly (exhibit “trance logic”). Finally, none of the 31 subjects whose handwriting was rated as “childlike and primitive” spelled the word “psychological” correctly, providing clear evidence that subjects who were the most responsive to the age regression suggestion fail to exhibit trance logic

Pheochromocytoma presenting as musculoskeletal pain from bone metastases

Six patients presented with musculoskeletal pain resulting from destructive bone lesions. These patients were ultimately shown to have metastatic pheochromocytoma. None of the cases exhibited typical symptoms of metastatic pheochromocytoma, nor was it suspected at the time of presentation. In three patients, hypertension caused pheochromocytoma to be considered as a diagnosis. The three remaining patients, all of whom had documented hypertension in the past, underwent bone biopsy. Two of these patients became markedly hypertensive in the postoperative period. Malignant pheochromocytoma may present with metastatic skeletal disease in some patients in whom the presence of hypertension as well as a carefully elicited history may suggest the diagnosis. In such patients, the possibility of pheochromocytoma should be taken into account, as biopsy may trigger a hypertensive crisis in patients not under adrenergic blockade.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/46787/1/256_2004_Article_BF00351270.pd

Distinct Pathways Mediate the Sorting of Tail-Anchored Proteins to the Plastid Outer Envelope

Background: Tail-anchored (TA) proteins are a distinct class of membrane proteins that are sorted post-translationally to various organelles and function in a number of important cellular processes, including redox reactions, vesicular trafficking and protein translocation. While the molecular targeting signals and pathways responsible for sorting TA proteins to their correct intracellular destinations in yeasts and mammals have begun to be characterized, relatively little is known about TA protein biogenesis in plant cells, especially for those sorted to the plastid outer envelope. Methodology/Principal Findings: Here we investigated the biogenesis of three plastid TA proteins, including the 33-kDa and 34-kDa GTPases of the translocon at the outer envelope of chloroplasts (Toc33 and Toc34) and a novel 9-kDa protein of unknown function that we define here as an outer envelope TA protein (OEP9). Using a combination of in vivo and in vitro assays we show that OEP9 utilizes a different sorting pathway than that used by Toc33 and Toc34. For instance, while all three TA proteins interact with the cytosolic OEP chaperone/receptor, AKR2A, the plastid targeting information within OEP9 is distinct from that within Toc33 and Toc34. Toc33 and Toc34 also appear to differ from OEP9 in that their insertion is dependent on themselves and the unique lipid composition of the plastid outer envelope. By contrast, the insertion of OEP9 into the plastid outer envelope occurs in a proteinaceous-dependent, but Toc33/34-independent manner and membrane lipids appear to serve primarily to facilitate normal thermodynamic integration of this TA protein. Conclusions/Significance: Collectively, the results provide evidence in support of at least two sorting pathways for plastid TA outer envelope proteins and shed light on not only the complex diversity of pathways involved in the targeting and insertion of proteins into plastids, but also the molecular mechanisms that underlie the delivery of TA proteins to their proper intracellular locations in general

Electrochemical Investigation of Azurin Thermodynamic and Adsorption Properties at Monolayer-Protected Cluster Film Assemblies – Evidence for a More Homogeneous Adsorption Interface

Thermodynamic and adsorption properties of protein monolayer electrochemistry (PME) are examined for Pseudomonas aeruginosa azurin (AZ) immobilized at an electrode modified with a networked film of monolayer-protected clusters (MPCs) to assess if nanoparticle films of this nature offer a more homogeneous adsorption interface compared to traditional self-assembled monolayer (SAM) modified electrodes. Specifically, electrochemistry is used to assess properties of surface coverage, formal potential, peak broadening, and electron transfer (ET) kinetics as a function of film thickness. The modification of a surface with dithiol-linked films of MPCs (Au225C675) provides a more uniform binding interface for AZ that results in voltammetry with less peak broadening (mV) compared to SAMs (\u3e120–130 mV). Improved homogeneity of the MPC interface for protein adsorption is confirmed by atomic force microscopy imaging that shows uniform coverage of the gold substrate topography and by electrochemical analysis of film properties during systematic desorption of AZ, which indicates a more homogeneous population of adsorbed protein at MPC films. These results suggest MPC film assemblies may be used in PME to provide greater molecular level control of the protein adsorption interface, a development with applications for strategies to study biological ET processes as well as the advancement of biosensor technologies

Investigating the Mechanisms of Hallucinogen-Induced Visions Using 3,4-Methylenedioxyamphetamine (MDA): A Randomized Controlled Trial in Humans

The mechanisms of drug-induced visions are poorly understood. Very few serotonergic hallucinogens have been studied in humans in decades, despite widespread use of these drugs and potential relevance of their mechanisms to hallucinations occurring in psychiatric and neurological disorders.We investigated the mechanisms of hallucinogen-induced visions by measuring the visual and perceptual effects of the hallucinogenic serotonin 5-HT2AR receptor agonist and monoamine releaser, 3,4-methylenedioxyamphetamine (MDA), in a double-blind placebo-controlled study. We found that MDA increased self-report measures of mystical-type experience and other hallucinogen-like effects, including reported visual alterations. MDA produced a significant increase in closed-eye visions (CEVs), with considerable individual variation. Magnitude of CEVs after MDA was associated with lower performance on measures of contour integration and object recognition.Drug-induced visions may have greater intensity in people with poor sensory or perceptual processing, suggesting common mechanisms with other hallucinatory syndromes. MDA is a potential tool to investigate mystical experiences and visual perception

Plume Characterization of a Laboratory Model 22 N GPIM Thruster via High-Frequency Raman Spectroscopy

The Green Propellant Infusion Mission (GPIM) will demonstrate the capability of a green propulsion system, specifically, one using the monopropellant, AF-M315E. One of the risks identified for GPIM is potential contamination of sensitive areas of the spacecraft from the effluents in the plumes of AF-M315E thrusters. Plume characterization of a laboratory-model 22 N thruster via optical diagnostics was conducted at NASA GRC in a space-simulated environment. A high-frequency pulsed laser was coupled with an electron-multiplied ICCD camera to perform Raman spectroscopy in the near-field, low-pressure plume. The Raman data yielded plume constituents and temperatures over a range of thruster chamber pressures and as a function of thruster (catalyst) operating time. Schlieren images of the near-field plume enabled calculation of plume velocities and revealed general plume structure of the otherwise invisible plume. The measured velocities are compared to those predicted by a two-dimensional, kinetic model. Trends in data and numerical results are presented from catalyst mid-life to end-of-life. The results of this investigation were coupled with the Raman and Schlieren data to provide an anchor for plume impingement analysis presented in a companion paper. The results of both analyses will be used to improve understanding of the nature of AF-M315E plumes and their impacts to GPIM and other future missions

The Green Propellant Infusion Mission Thruster Performance Testing for Plume Diagnostics

The Green Propellant Infusion Mission (GPIM) is sponsored by NASA's Space Technology Mission Directorate (STMD) Technology Demonstration Mission (TDM) office. The goal of GPIM is to advance the technology readiness level of a green propulsion system, specifically, one using the monopropellant, AF-M315E, by demonstrating ground handling, spacecraft processing, and on-orbit operations. One of the risks identified for GPIM is potential contamination of sensitive spacecraft surfaces from the effluents in the plumes of AF-M315E thrusters. NASA Glenn Research Center (GRC) is conducting activities to characterize the effects of AF-M315E plume impingement and deposition. GRC has established individual plume models of the 22-N and 1-N thrusters that will be used on the GPIM spacecraft. The model simulations will be correlated with plume measurement data from Laboratory and Engineering Model 22-N, AF-M315E thrusters. The thrusters are currently being tested in a small rocket, altitude facility at NASA GRC. A suite of diagnostics, including Raman spectroscopy, Rayleigh spectroscopy, and Schlieren imaging are being used to acquire plume measurements of AF-M315E thrusters. Plume data will include temperature, velocity, relative density, and species concentration. The plume measurement data will be compared to the corresponding simulations of the plume model. The GRC effort will establish a data set of AF-M315E plume measurements and a plume model that can be used for future AF-M315E applications