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M R Walter

Publications and source records attributed to M R Walter.

34 records · Page 2Linked to original sources

Purification and crystallization of a complex between human interferon gamma receptor (extracellular domain) and human interferon gamma.

X-ray diffraction quality crystals have been obtained from a complex between interferon gamma and the extracellular domain of its high-affinity cell surface receptor. The crystals were obtained from interferon gamma/interferon gamma receptor complexes purified by size exclusion chromatography. Diffraction quality crystals required analyzing these complex samples by isoelectric focusing gels to select purified complex fractions devoid of unbound interferon gamma. These studies used interferon gamma receptor engineered with an eight amino acid N-terminal deletion to eliminate heterogeneity generated due to proteolytic cleavage. In addition, the receptor was expressed in an E. coli secretion cell line which eliminated the need to refold the protein. Hexagonal crystals were grown from 1.6 M ammonium phosphate solutions and belong to a spacegroup of P6(5)22 with unit cell dimensions a = 145.9 A and c = 180.3 A. These crystals diffract to at least 2.9 A resolution when exposed to synchrotron radiation. SDS PAGE analysis of the crystals demonstrated that both interferon gamma and the receptor were present. Analysis of the x-ray diffraction data revealed that the crystals contain complexes with a stoichiometry of 2:1 receptor: ligand within the crystallographic asymmetric unit and consist of approximately 55% solvent.

Chromatography, Gel↗

The limits of palaeontological knowledge: finding the gold among the dross.

Palaeontological interpretation rests on two interwoven sets of comparisons with the modern world. Palaeobiological interpretation relies on the placement of fossils within a phylogenetic and functional framework based primarily on the comparative biology of living organisms. Analogy to currently observable chemical, physical and taphonomic processes enables palaeoenvironmental inferences to be drawn from geological data. In older rocks, comparisons with the modern Earth can become tenuous, limiting palaeontological interpretation. The problem reaches its apogee in Archaean successions, yet pursuit of multiple lines of evidence establishes that complex microbial communities, fuelled by autotrophy and, likely, photoautotrophy, existed 3500 million years ago. Although Archaean palaeontology has to date focused on silicified coastal sediments, improved understanding of Earth's earliest biosphere may depend on the development of alternative environmental and taphonomic analogies. Spring precipitates and hydrothermal metal deposits are promising candidates. Terrestrial organisms may be of limited value in interpreting such fossils as may be found on Mars, although some points of comparison could prove general. Given limited opportunities for exploration, proper choice of environmental analogy is critical. Spring precipitates constitute excellent deposits for addressing questions of biology on another planet.

Archaea↗

Ancient hydrothermal ecosystems on earth: a new palaeobiological frontier.

Thermal springs are common in the oceans and on land. Early in the history of the Earth they would have been even more abundant, because of a higher heat flow. A thermophilic lifestyle has been proposed for the common ancestor of extant life, and hydrothermal ecosystems can be expected to have existed on Earth since life arose. Though there has been a great deal of recent research on this topic by biologists, palaeobiologists have done little to explore ancient high temperature environments. Exploration geologists and miners have long known the importance of hydrothermal systems, as they are sources for much of our gold, silver, copper, lead and zinc. Such systems are particularly abundant in Archaean and Proterozoic successions. Despite the rarity of systematic searches of these by palaeobiologists, already 12 fossiliferous Phanerozoic deposits are known. Five are 'black smoker' type submarine deposits that formed in the deep ocean and preserve a vent fauna like that in the modern oceans; the oldest is Devonian. Three are from shallow marine deposits of Carboniferous age. As well as 'worm tubes', several of these contain morphological or isotopic evidence of microbial life. The oldest well established fossiliferous submarine thermal spring deposit is Cambro-Ordovician; microorganisms of at least three or four types are preserved in this. One example each of Carboniferous and Jurassic sub-lacustrine fossiliferous thermal springs are known. There are two convincing examples of fossiliferous subaerial hydrothermal deposits. Both are Devonian. Several known Proterozoic and Archaean deposits are likely to preserve a substantial palaeobiological record, and all the indications are that there must be numerous deposits suitable for study. Already it is demonstrable that in ancient thermal spring deposits there is a record of microbial communities preserved as stromatolites, microfossils, isotope distribution patterns and hydrocarbon biomarkers.

Animals↗

Lithofacies and biofacies of mid-Paleozoic thermal spring deposits in the Drummond Basin, Queensland, Australia.

The Devonian to Carboniferous sinters of the Drummond Basin, Australia, are among the oldest well established examples of fossil subaerial hot springs. Numerous subaerial and subaqueous spring deposits are known from the geological record as a result of the occurrence of economic mineral deposits in many of them. Some are reported to contain fossils, but very few have been studied by paleobiologists; they represent an untapped source of paleobiological information on the history of hydrothermal ecosystems. Such systems are of special interest, given the molecular biological evidence that thermophilic bacteria lie near the root of the tree of extant life. The Drummond Basin sinters are very closely comparable with modern examples in Yellowstone National Park and elsewhere. Thirteen microfacies are recognisable in the field, ranging from high temperature apparently abiotic geyserite through various forms of stromatolitic sinter probably of cyanobacterial origin to ambient temperature marsh deposits. Microfossils in the stromatolites are interpreted as cyanobacterial sheaths. Herbaceous lycopsids occur in the lower temperature deposits.

Australia↗

Crystal structure of interleukin 10 reveals an interferon gamma-like fold.

The crystal structure of recombinant human interleukin 10 (rhIL-10) has been determined by X-ray crystallography at 2.0 A resolution. Interleukin 10 is a dimer composed of identical polypeptide chains related by a 2-fold axis. The molecule is predominantly alpha-helical. The main-chain fold resembles that of interferon gamma (IFN-gamma) in which the structural integrity of each domain is dependent on the intertwining of helices from each peptide chain. Comparison of rhIL-10 and IFN-gamma reveals differences in helix lengths and orientations of the 2-fold related domains. Interleukin 10 and IFN-gamma contain several conserved residues in their internal cores which suggest a possible "fingerprint" for detection of other members of this fold.

Amino Acid Sequence↗

Crystal structure of a complex between interferon-gamma and its soluble high-affinity receptor.

The crystal structure of interferon-gamma bound to the extracellular fragment of its high-affinity cell-surface receptor reveals the first view of a class-2 cytokine receptor-ligand complex. In the complex, one interferon-gamma homodimer binds two receptor molecules. Unlike the class-1 growth hormone receptor complex, the two interferon-gamma receptors do not interact with one another and are separated by 27 A. Upon receptor binding, the flexible AB loop of interferon-gamma undergoes a conformational change that includes the formation of a 3(10) helix.

Computer Graphics↗

Crystallization and preliminary X-ray investigation of recombinant human interleukin 10.

Crystals of recombinant human interleukin 10 have been grown from solutions of ammonium sulfate. The crystals are tetragonal, space group P4(1)2(1)2 or P4(3)2(1)2; the unit cell axes are a = 36.5 A and c = 221.9 A. There is the equivalent of one polypeptide chain in the asymmetric unit. The crystals are stable to X-rays and diffract to at least 2.5 A resolution.

Ammonium Sulfate↗

Drug binding by calmodulin: crystal structure of a calmodulin-trifluoperazine complex.

The crystal structure of calmodulin (CaM) bound to trifluoperazine (TFP) has been determined and refined to a resolution of 2.45 A. Only one TFP is bound to CaM, but that is sufficient to cause distortion of the central alpha-helix and juxtaposition of the N- and C-terminal domains similar to that seen in CaM-polypeptide complexes. The drug makes extensive contacts with residues in the C-terminal domain of CaM but only a few contacts with one residue in the N-terminal domain. The structure suggests that substrate binding to the C-terminal domain is sufficient to cause the conformational changes in calmodulin that lead to activation of its targets.

Animals↗

Comparison of four independently determined structures of human recombinant interleukin-4.

Four independent structures of human interleukin-4, two determined by nuclear magnetic resonance techniques and two by X-ray diffraction, have been compared in detail. The core of this four helix bundle protein is very similar in all the structures but there are some differences in loop regions that are known to be mobile in solution. Careful comparison of the experimental data sets and the methods of analysis of the different laboratories has provided clues to the sources of most of the differences, and also answered some general questions about the accuracy of protein structure determination by these two techniques.

Crystallography, X-Ray↗

Preservation of biological information in thermal spring deposits: developing a strategy for the search for fossil life on Mars.

Current interpretations of the early history of Mars suggest many similarities with the early Earth and therefore raise the possibility that the Archean and Proterozoic history of life on Earth could have a counterpart on Mars. Terrestrial experience suggests that, with techniques that can be employed remotely, ancient springs, including thermal springs, could well yield important information. By delivering water and various dissolved species to the sunlit surface of Mars, springs very likely created an environment suitable for life, which could have been difficult, if not impossible, to attain elsewhere. The chemical and temperature gradients associated with thermal springs sort organisms into sharply delineated, distinctive and different communities, and so diverse organisms are concentrated into relatively small areas in a predictable and informative fashion. A wide range of metabolic strategies are concentrated into small areas, thus furnishing a useful and representative sampling of the existing biota. Mineral-charged springwaters frequently deposit chemical precipitates of silica and/or carbonate which incorporate microorganisms and preserve them as fossils. The juxtaposition of stream valley headwaters with volcanoes and impact craters on Mars strongly implies that subsurface heating of groundwater created thermal springs. On Earth, thermal springs create distinctive geomorphic features and chemical signatures which can be detected by remote sensing. Spring deposits can be quite different chemically from adjacent rocks. Individual springs can be hundreds of meters wide, and complexes of springs occupy areas up to several kilometers wide. Benthic microbial mats and the resultant stromatolites occupy a large fraction of the available area. The relatively high densities of fossils and microbial mat fabrics within these deposits make them highly prospective in any search for morphological evidence of life, and there are examples of microbial fossils in spring deposits as old as 300 Myr.

Bacteria↗

Crystal structure of recombinant human interleukin-4.

The crystal structure of recombinant human interleukin-4 (rhuIL-4) was initially determined at 3.5-A resolution by multiple isomorphous replacement techniques and subsequently refined to a resolution of 2.35 A by simulated annealing. The final crystallographic R-factor, based on all data in the range 6.0-2.35 A (7470 reflections), is 0.232. Bond lengths and bond angles in the molecule have root mean square deviations from ideal values of 0.016 A and 2.4 degrees, respectively. The overall structure is highly compact and globular with a predominantly hydrophobic core. The main structural feature of rhuIL-4 is a four alpha-helix bundle, which composes approximately 58% of the structure. The helices are arranged in a left-handed antiparallel bundle with two overhand connections. Within these connections is a two-stranded antiparallel beta-sheet. Both the tertiary and secondary structures of rhuIL-4 are similar to those of human granulocyte-macrophage colony-stimulating factor. Critical regions for receptor binding are proposed.

Amino Acid Sequence↗

Latest Proterozoic stratigraphy and Earth history.

The end of the Proterozoic Eon was a time of pronounced biological, biogeochemical, climatic and tectonic change. New bio- and chemostratigraphic data provide an improved framework for stratigraphic correlation, making possible a deeper understanding of latest Proterozoic Earth history and providing tools for a chronostratigraphic division of late Proterozoic time.

Animals↗

Three-dimensional structure of recombinant human granulocyte-macrophage colony-stimulating factor.

The crystal structure of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) has been determined at 2.8 A resolution using multiple isomorphous replacement techniques. There are two molecules in the crystallographic asymmetric unit, which are related by an approximate non-crystallographic 2-fold axis. The overall structure is highly compact and globular with a predominantly hydrophobic core. The main structural feature of rhGM-CSF is a four alpha-helix bundle, which represents approximately 42% of the structure. The helices are arranged in a left-handed antiparallel bundle with two overhand connections. Within the connections is a two-stranded antiparallel beta-sheet. The tertiary structure of rhGM-CSF has a topology similar to that of porcine growth factor and interferon-beta. Most of the proposed critical regions for receptor binding are located on a continuous surface at one end of the molecule that includes the C terminus.

Amino Acid Sequence↗

Three-dimensional structure of thymidine phosphorylase from Escherichia coli at 2.8 A resolution.

The three-dimensional structure of thymidine phosphorylase from Escherichia coli has been determined at 2.8 A resolution using multiple-isomorphous-replacement techniques. The amino acid sequence deduced from the deoA DNA sequence is also reported. Thymidine phosphorylase exists in the crystal as an S-shaped dimer in which the subunits are related by a crystallographic 2-fold axis. Each subunit is composed of a small alpha-helical domain of six helices and a large alpha/beta domain. The alpha/beta domain includes a six-stranded mixed beta-sheet and a four-stranded antiparallel beta-sheet. The active site has been identified by difference Fourier analyses of the binding of thymine and thymidine and lies in a cavity between the small and large domains. The central beta-sheet is splayed open to accommodate a putative phosphate-binding site which is probably occupied by a sulfate ion in the crystal.

Amino Acid Sequence↗

Millimeter-scale variations of stable isotope abundances in carbonates from banded iron-formations in the Hamersley Group of Western Australia.

Several diamond drill cores from formations within the Hamersley Group of Western Australia have been studied for evidence of short-range variations in the isotopic compositions of the carbonates. For a set of 32 adjacent microbands analyzed in a specimen from the Marra Mamba Iron Formation, carbon isotope compositions of individual microbands ranged from -2.8 to -19.8 per mil compared to PDB and oxygen isotope compositions ranged from 10.2 to 20.8 per mil compared to SMOW. A pattern of alternating abundances was present, with the average isotopic contrasts between adjacent microbands being 3.0 per mil for carbon and 3.1 per mil for oxygen. Similar results were obtained for a suite of 34 microbands (in four groups) from the Bruno's Band unit of the Mount Sylvia Formation. Difficulties were experienced in preparing samples of single microbands from the Dales Gorge Member of the Brockman Iron Formation, but overall isotopic compositions were in good agreement with values reported by previous authors. Chemical analyses showed that isotopically light carbon and oxygen were correlated with increased concentrations of iron. The preservation of these millimeter-scale variations in isotopic abundances is interpreted as inconsistent with a metamorphic origin for the isotopically light carbon in the BIF carbonates. A biological origin is favored for the correlated variations in 13C and Fe, and it is suggested that the 13C-depleted carbonates may derive either from fermentative metabolism or from anaerobic respiration. A model is presented in which these processes occur near the sediment-water interface and are coupled with an initial oxidative precipitation of the iron.

Australia↗