Engineering of zinc finger and MHC motifs to locked-in tertiary folds

The assembly of helical and -sheet peptide blocks containing reactive chain ends results inhighly branched chain architectures (locked-in folds) mimicking native tertiary structures.This molecular kit strategy allows to bypass the protein folding problem in protein de novodesign and gives access to protein mimetics of high thermodynamic stability. The validity ofthis concept is exemplified for the design and synthesis of locked-in folds mimicking the zincfinger and MHC folding motifs.     

Molecular cloning of a new receptor-like kinase gene encoded at the Lr10 disease resistance locus of wheat

More than 100 resistance genes against wheat rust pathogens have been described in wheat and its relatives. Although many of them have been extensively used in wheat resistance breeding, none of these resistance loci has yet been analyzed at the molecular level. By screening a set of near-isogenic lines carrying different leaf rust resistance genes with a wheat probe encoding a serine/threonine protein kinase, we detected a polymorphic DNA fragment in the line with the Lr10 resistance gene. This fragment mapped to the Lr10 disease resistance locus and encodes a receptor-like protein kinase which we called LRK10. LRK10 contains a new type of extracellular domain not found in known plant or animal receptor kinases. Several conserved amino acids in S-domain glycoproteins and receptor-like kinases were also found in LRK10, suggesting that LRK10 and S-domain proteins belong to the same superfamily of specific recognition molecules in plants. Lrk10 was expressed at low levels in young seedlings and belongs to a gene family. Analysis of wheat lines with and without the Lr10 gene demonstrated that Lrk10 and Lr10 belong to the same genetic locus. We conclude that gene isolation based on protein kinase homology can identify new receptor domains and provide candidates for disease resistance genes in the complex wheat genome.     

Immunological studies on lysosomal sphingomyelinase: Identification of a 28 000-Da component deficient in urine from patients with Niemann-Pick disease types A and B

The immunoblotting technique was used to identify sphingomyeJinase protein in samples of tissue and urine after subjection to poIyacrylamide-gel etectrophoresis in the presence of sodium dodecyl sulphate. In a sphingomyelinase preparation purified from control urine a prominent band was seen with an M_r of 28 000 Da. Glycoprotein fractions from urine and placenta, a membrane extract from spleen, and a partially purified sphingomyelinase preparation from placenta contained the 28 000-Da band plus additional, higher-M_r bands. The 28 000-Da band was detectable in urine from a patient with Niemann-Pick disease type C, but not in urine from patients with Niemann-Pick disease types A and B. It is concluded t h a t sphingomyeJinase is composed of at least one polypeptide with an M_r of 28 000 Da and that this polypeptide is deficient in the urine of patients with Niemann-Pick disease types A and B.     

Nickel requirement in Chlorella emersonii

The dependence of growth on nickel supply was studied in Chlorella emersonii 211-8b. After transfer to Ni²⁺ deficient medium containing only 0.5±0.2 g/l of Ni²⁺, production of biomass or daughter cells dropped to 55±5% of the controls, and the cells became chlorotic. These symptoms of deficiency disappeared completely by supplying adequate amounts of nickel. They were, however, only partially reversible by cobalt. It is concluded that nickel is an essential micronutrient for C. emersonii, although this organism lacks the nickel enzyme, urease.Gratefully dedicated to Prof. Hans Adolf von Stosch on the occasion of his 70th birthday     

Oceanographic significance of Pacific late miocene calcareous nannoplankton

An analysis of the variability in the composition and distribution of Pacific Late Miocene calcareous nannoplankton about their average biogeography shows that there are primarily two environmental factors causing that variability, climate and dissolution. Climate produces a latitudinal, biogeographic differentiation of the Late Miocene nannoflora, while selective dissolution superimposes a bathymetric differentiation of the nannoflora on that due to climate. Together, these two factors produce three distinct Late Miocene nannofloral assemblages, a high-latitude, temperate assemblage characterized by Reticulofenestra pseudoumbilica and Coccolithus pelagicus, and two tropical assemblages, their differences in composition depending on water depth and surface-water productivity: (1) in shallower water and beneath areas of higher organic production and sedimentation of calcite there is an undissolved assemblage characterized by sphenoliths, small elliptical placoliths and Coccolithus pataecus; (2) in deeper water and areas of lower productivity there is a dissolved assemblage dominated by discoasters.Selective dissolution produces most of the apparent biogeographic variation in Pacific Late Miocene nannoplankton compositions, the variation in compositions observed between the seventeen sites studied. Dissolution preferentially removes the more soluble constituents of the tropical nannoflora so that increasing dissolution tends to give tropical nannoflora a cooler, more temperate aspect. At the same time, selective dissolution shifts the composition of the warmer, tropical component towards its more resistant taxa.Nannoplankton records show a period of greatly decreased calcite dissolution in deep tropical and temperate South Pacific sites between about 8 and 10 m.y. ago. This decrease is strongly correlated with a temporary increase in the ¹³C composition of Pacific deep waters. Calcite dissolution increased during this same period in the deep North Pacific.Nannoplankton records of Late Miocene climate in the tropics are distinctly different from those at higher, south temperate latitudes. Tropical records show a sharp warming in the earliest Late Miocene after a generally cool late Middle Miocene. This was followed by a temporary cooling, nearly to Middle Miocene levels, about 7 m.y. ago. Toward the end of the Late Miocene, the tropical Pacific warmed again and remained warm into the Pliocene. Warming of temperate climates occurred much later. Not until latest Miocene did the southern the Pliocene. Warming of temperate climates occurred much later. Not until latest Miocene did the southern temperate latitudes warm appreciably. Southern subpolar climate cooled continuously through the Late Miocene. We attribute the resulting increases in the latitudinal climatic contrast across the southern Pacific Ocean to the development and migration of a strong subtropical convergence.On the basis of the nannoplankton oceanographic records we postulate that beginning about 10.5 m.y. ago Pacific surface circulation became primarily zonal and the production of deep and bottom waters in the Southern Ocean increased sharply. This produced a northward decrease in calcite preservation, an increase in benthic ¹³C, and a strong climatic gradient across southern latitudes. The period of most vigorous deep Pacific circulation ended 7 m.y. ago in response, we speculate, to the reduced ocean salinities during the Messinian.     

Degradation of Coal by the Fungi Polyporus versicolor and Poria monticola

We report that two species of basidiomycete fungi (Polyporus versicolor and Poria monticola) grow in minimal liquid or solid medium when supplemented with crushed lignite coal. The fungi also grow directly on crushed lignite coal. The growth of both fungi was observed qualitatively as the production and extension of hyphae. No fungal growth occurred in minimal agar medium without coal. The fungi degraded solid lignite coal to a black liquid product which never appeared in cultures unless fungi and coal were present together. Apparently, lignite coal can serve as the principal substrate for the growth of the fungi. Infrared analyses of the liquid products of lignite degradation showed both similarities to and differences from the original lignite.     

Chick embryo muscarinic and purinergic receptors activate cytosolic Ca2+ via phosphatidylinositol metabolism

In dissociated cells from chick embryos or from chick limb buds, acetylcholine (ACh) induced an increase in cellular levels of inositol 1,4,5-trisphosphate (Ins-P3) and of inositol 1,3,4,5-tetrakisphosphate (Ins-P4). The concentration of Ins-P3 was enhanced transiently, whereas the level of Ins-P4 remained elevated for at least 20 min after addition of ACh. In most cases the increase in Ins-P4 levels was more pronounced than that of Ins-P3 levels. The inhibition of the ACh-induced inositol-phosphate response by atropine (half-maximal inhibition at 10 nM) indicates the involvement of muscarinic receptors, which in chick embryo cells induce a transient rise and a following persistent elevation of cytosolic Ca2+ activity (G. Oettling et al. (1989) J. Dev. Physiol. 12, 85-94). Adenosine 5'-triphosphate (ATP) elicited a similar transient rise in cytosolic Ca2+ activity, however, without a subsequent plateau. ATP also caused an increase in inositol-oligophosphate levels. Thus, both muscarinic and purinergic receptors in chick embryo cells are coupled to phospholipase C. The enzymatically formed Ins-P3 mediates the release of Ca2+ from internal stores. The Ca2+ signal could be involved in embryonic cell migration during morphogenesis.