Crystallization of insecticyanin from the hemolymph of the tobacco hornworm Manduca sexta L. in a form suitable for a high resolution structure determination

Insecticyanin, a blue biliprotein from the tobacco hornworm Manduca sexta, has been crystallized in a form suitable for a high resolution x-ray analysis. The crystals grow by vapor diffusion against solutions of polyethylene glycol 8000 at pH 5.5. They belong to the space group P4(1)2(1)2 or P4(3)2(1)2 with unit cell dimensions of a = b = 115.0 A; c = 71.1 A. Insecticyanin is believed to be a tetramer in solution; there are two subunits per asymmetric unit. The crystals diffract to at least 2.2 A resolution and appear reasonably resistant to radiation damage.     

Myosin subfragment-1: structure and function of a molecular motor: Current opinion in structural biology 1993, 3:944–952

The ability for directed movement is a fundamental process of all living systems. Molecules designed for such purposes are ubiquitous in eukaryotic cells and have been the focus of intense investigations for many years. Highlighted in this report is the three-dimensional structure of the myosin motor domain—the first such motor protein to be examined by single-crystal X-ray diffraction analysis.     

Three-dimensional structure of myosin subfragment-1 from electron microscopy of sectioned crystals

Image analysis of electron micrographs of thin-sectioned myosin subfragment-1 (S1) crystals has been used to determine the structure of the myosin head at approximately 25-A resolution. Previous work established that the unit cell of type I crystals of myosin S1 contains eight molecules arranged with orthorhombic space group symmetry P212121 and provided preliminary information on the size and shape of the myosin head (Winkelmann, D. A., H. Mekeel, and I. Rayment. 1985. J. Mol. Biol. 181:487-501). We have applied a systematic method of data collection by electron microscopy to reconstruct the three-dimensional (3D) structure of the S1 crystal lattice. Electron micrographs of thin sections were recorded at angles of up to 50 degrees by tilting the sections about the two orthogonal unit cell axes in sections cut perpendicular to the three major crystallographic axes. The data from six separate tilt series were merged to form a complete data set for 3D reconstruction. This approach has yielded an electron density map of the unit cell of the S1 crystals of sufficient detail. to delineate the molecular envelope of the myosin head. Myosin S1 has a tadpole-shaped molecular envelope that is very similar in appearance to the pear-shaped myosin heads observed by electron microscopy of rotary-shadowed and negatively stained myosin. The molecule is divided into essentially three morphological domains: a large domain on one end of the molecule corresponding to approximately 60% of the total molecular volume, a smaller central domain of approximately 30% of the volume that is separated from the larger domain by a cleft on one side of the molecule, and the smallest domain corresponding to a thin tail-like region containing approximately 10% of the volume. This molecular organization supports models of force generation by myosin which invoke conformational mobility at interdomain junctions within the head.     

Crystallization and preliminary analysis of telokin, the C-terminal domain of myosin light chain kinase

Telokin, an acidic protein related to the C-terminal portion of smooth muscle myosin light chain kinase from turkey gizzard has been crystallized in a form suitable for a high-resolution diffraction analysis. The crystals were grown from solutions of polyethylene glycol 8000 using the hanging-drop vapor diffusion method. They belong to the trigonal space group P3(1)21 or P3(2)21 with cell parameters a = 64.0 A, c = 59.4 A and diffract to at least 2.7 A resolution.     

Trimethyllead acetate: a first-choice heavy atom derivative for protein crystallography

The three-dimensional conformation of a protein provides a wealth of biochemical information and with the advent of cloning techniques that allow the preparation of proteins almost at will, a renewed interest has arisen in the crystallographic determination of protein structures. As in any research technique, however, there are often many difficulties encountered in an X-ray crystallographic investigation. One of these is the "phase problem." Although in recent years there has been considerable progress in the development of techniques for phase determination, including the use of molecular replacement and multiple wavelength measurements, the multiple isomorphous replacement method is still the most successful method for obtaining a three-dimensional structure. Here we report the use of trimethyllead acetate as a heavy atom compound of first choice in the preparation of an isomorphous heavy atom derivative.     

Uncertainty in research about key invasion characteristics limits the evaluation of exotic perennial grasses in natural systems in New South Wales,Australia

Exotic perennial grasses (EPGs) pose a significant risk to native communities globally. With over 2,200 species in Australia, understanding which characteristics enable high threat invasions, and comparing between functionally similar EPGs, can help prioritise species management. We developed a framework of risk and used the literature to rank 21 EPGs considered a threat to plant communities in New South Wales, while also evaluating the reliability of information currently available. Characteristics were scored within five broad categories that distinguish invasiveness: Arrival, Establishment, Persistence, Impact and Distribution. These included aspects of reproductive biology, competitive ability and environmental tolerance. The risk assessment was effective in assessing key characteristics of invasion. EPGs with an economic benefit (trade‐off species) were more likely to have reliable research and frequently ranked as high‐risk invaders in natural habitats due to the overlap of characteristics important in invasion with those considered important in agriculture. Lack of formal scientific research hindered assessment for some species, and some traits had been poorly assessed in the literature. High uncertainty was associated with key characteristics for Establishment, Persistence and Impact. Uncertainty in key characteristics revealed a need for improved integration of less formal research validated by more formal scientific research. This may lead to more informed decisions in the management of EPGs in native habitats and assist in early control of EPGs not yet assessed.     

Bacterial composition of soils of the Lake Wellman area, Darwin Mountains, Antarctica

The aim of this study was to examine the bacterial composition of high latitude soils from the Darwin–Hatherton glacier region of Antarctica. Four soil pits on each of four glacial drift sheets were sampled for chemical and microbial analyses. The four drifts—Hatherton, Britannia, Danum, and Isca—ranged, respectively, from early Holocene (10 ky) to mid-Quaternary (ca 900 ky). Numbers of culturable bacteria were low, with highest levels detected in soils from the younger Hatherton drift. DNA was extracted and 16S rRNA gene clone libraries prepared from samples below the desert pavement for each of the four drift sheets. Between 31 and 262 clones were analysed from each of the Hatherton, Britannia, and Danum drifts. Bacterial sequences were dominated by members of the phyla Deinococcus-Thermus, Actinobacteria, and Bacteroidetes. Culturable bacteria, including some that clustered with soil clones (e.g., members of the genera Arthrobacter, Adhaeribacter, and Pontibacter), belonged to Actinobacteria and Bacteroidetes. The isolated bacteria are ideal model organisms for genomic and phenotypic investigations of those attributes that allow bacteria to survive and/or grow in Antarctic soils because they have close relatives that are not tolerant of these conditions.     

Spatial variation in the ongoing and widespread decline of a keystone plant species

Extensive dieback in dominant plant species in response to climate change is increasingly common. Climatic conditions and related variables, such as evapotranspiration, vary in response to topographical complexity. This complexity plays an important role in the provision of climate refugia. In 2008/2009, an island‐wide dieback event of the keystone cushion plant Azorella macquariensis Orchard (Apiaceae) occurred on sub‐Antarctic Macquarie Island. This signalled the start of a potential regime shift, suggested to be driven by increasing vapour pressure deficit. Eight years later, we quantified cover and dieback across the range of putative microclimates to which the species is exposed, with the aim of explaining dieback patterns. We test for the influence of evapotranspiration using a suite of topographic proxies and other variables as proposed drivers of change. We found higher cover and lower dieback towards the south of the island. The high spatial variation in A. macquariensis populations was best explained by latitude, likely a proxy for macroscale climate gradients and geology. Dieback was best explained by A. macquariensis cover and latitude, increasing with cover and towards the north of the island. The effect sizes of terrain variables that influence evapotranspiration rates were small. Island‐wide dieback remains conspicuous. Comparison between a subset of sites and historical data revealed a reduction of cover in the north and central regions of the island, and a shift south in the most active areas of dieback. Dieback remained comparatively low in the south. The presence of seedlings was independent of dieback. This study provides an empirical baseline for spatial variation in the cover and condition of A. macquariensis, both key variables for monitoring condition and ‘cover‐debt’ in this critically endangered endemic plant species. These findings have broader implications for understanding the responses of fellfield ecosystems and other Azorella species across the sub‐Antarctic under future climates.