Multiple conformations of a protein demonstrated by magnetization transfer NMR spectroscopy

It is generally accepted that a globular protein in its native state adopts a single, well-defined conformation. However, there have been several reports that some proteins may exist in more than one distinct folded form in equilibrium. In the case of staphylococcal nuclease, evidence for multiple conformations has come from electrophoretic and NMR studies, although there has been some controversy as to whether these are actually interconvertible forms of the same molecular species. Recently, magnetization transfer (MT)-NMR has been developed as a means of studying the kinetics of conformational transitions in proteins. In the study reported here, this approach has been extended and used to demonstrate the presence of at least two native forms of nuclease in equilibrium and to study their interconversion with the unfolded state under the conditions of the thermal unfolding transition. The experiments reveal that two distinct native forms of the protein fold and unfold independently and that these can interconvert directly as well as via the unfolded state. The spectra of the different forms suggest that they are structurally similar but the MT experiments show that the kinetics of folding and unfolding are quite different. Characterization of this behaviour will, therefore, have important implications for our understanding of the relationship between structure and folding kinetics.     

Excitatory amino acids inhibit stimulation of phosphatidylinositol metabolism by aminergic agonists in hippocampus

Since the initial observations in the 1950s a large number of neurotransmitters and hormones have been shown to influence phosphatidylinositol (PI) metabolism in brain and peripheral ganglia (see ref. 3 for review). This has led to the suggestion that PI is part of an intracellular second messenger system for some types of diffusible chemical factors. Consistent with this are recent reports that one of the products of PI turnover (diacylglycerol) stimulates the Ca-dependent phospholipid-dependent protein kinase (kinase C) while a second (inositol trisphosphate) causes the release of calcium from intracellular stores. Thus it is possible that at least some brain neurotransmitters utilize the PI system to produce functional effects that are in addition to and which outlast the very brief physiological responses they elicit. Although it had been anticipated that another class of receptors might inhibit receptor-mediated stimulation of PI breakdown, no clear examples of such effects have been described. We now report that acidic amino acids, which are that acidic amino acids, which are thought to be excitatory neurotransmitters at the majority of brain synapses, strongly inhibit the stimulation of PI metabolism elicited by carbachol, histamine, or by potassium-induced depolarization, without changing the response to noradrenaline. As well as indicating a novel function for the excitatory amino acids, these results suggest that the central nervous system possesses cell-cell interactions of a previously unsuspected type.     

Protein encoded by v-erbA functions as a thyroid-hormone receptor antagonist

The thyroid-hormone receptor can, in the absence of its ligand, suppress activity of a responsive promoter. Addition of thyroid hormone, however, results in the stimulation of expression. The oncogenic derivative of the thyroid-hormone receptor, v-erbA, acts as a constitutive repressor and, when coexpressed with the receptor, blocks activation by thyroid hormone. Thus, v-erbA may be the first example of a dominant negative oncogene.     

Scanning tunnelling microscopy of Z-DNA

Scanning tunnelling microscopy (STM) has been used to map the surface topography of inorganic materials at the atomic level, and is potentially one of the most powerful techniques for probing biomolecular structure. Recent STM studies of calf thymus DNA and poly(rA).poly(rU) have shown that the helical pitch and periodic alternation of major and minor grooves can be visualized and reliably measured. Here we present the first STM images of poly(dG-me5dC).poly(dG-me5dC) in the Z-form. Both the general appearance of the fibres and measurements of helical parameters are in good agreement with models derived from X-ray diffraction.     

Identification of the drug Darvon and its metabolites in the urine of a comatose patient using a gas chromatograph-mass. Spectrometer-computer system

Zusammenfassung Mit Hilfe von hoch- und niederauflösender Massenspektroskopie sowie Daten eines Gas-chromatograph-Massenspektrometer-Computer-Systems wurden die Droge Darvon³ sowie eine Reihe ihrer Metaboliten im Harn eines Überdosispatienten nachgewiesen.

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This publication describes the identification of a drug causing coma in a patient. The drug was identified by analysis of the patient's urine performed at the NIH sponsored Special Research Resource Facility for Mass Spectrometry at M.I.T., under emergency conditions. Most of the experiments, measurements and interpretation reported had to be made within 1 day, thus requiring the simultaneous efforts of many members of M.I.T.'s massspectrometry group.     

A rare type of ascidium

Résumé Un type d'ascidie peu commun et jusqu'ici non enregistré dans les plantes charnues est décrit. Le point de croissance à l'intérieur du gobelet n'avorta pas, de sorte que le rejeton continua à croître à partir de l'intérieur de l'ascidie pour aboutir à une tête florale.     

Synthetic GPI as a candidate anti-toxic vaccine in a model of malaria

The malaria parasite Plasmodium falciparum infects 5-10% of the world's population and kills two million people annually. Fatalities are thought to result in part from pathological reactions initiated by a malarial toxin. Glycosylphosphatidylinositol (GPI) originating from the parasite has the properties predicted of a toxin; however, a requirement for toxins in general and GPI in particular in malarial pathogenesis and fatality remains unproven. As anti-toxic vaccines can be highly effective public health tools, we sought to determine whether anti-GPI vaccination could prevent pathology and fatalities in the Plasmodium berghei/rodent model of severe malaria. The P. falciparum GPI glycan of the sequence NH(2)-CH(2)-CH(2)-PO(4)-(Man alpha 1-2)6Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH(2)alpha 1-6myo-inositol-1,2-cyclic-phosphate was chemically synthesized, conjugated to carriers, and used to immunize mice. Recipients were substantially protected against malarial acidosis, pulmonary oedema, cerebral syndrome and fatality. Anti-GPI antibodies neutralized pro-inflammatory activity by P. falciparum in vitro. Thus, we show that GPI is a significant pro-inflammatory endotoxin of parasitic origin, and that several disease parameters in malarious mice are toxin-dependent. GPI may contribute to pathogenesis and fatalities in humans. Synthetic GPI is therefore a prototype carbohydrate anti-toxic vaccine against malaria.