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Now showing items 1 - 16 of 390

  • Ancestral protein resurrection and engineering opportunities of the mamba aminergic toxins

    Guillaume Blanchet   Enrico A. Stura   Gilles Mourier   Nicolas Gilles   Denis Servent  

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  • Crystal Structure of Human Prostate-Specific Antigen in a Sandwich Antibody Complex

    Enrico A. Stura   Bruno H. Muller   Marc Bossus   Sandrine Michel   Colette Jolivet-Reynaud   Frédéric Ducancel  

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  • Small molecule mimetics of erythropoietin

    The invention features computer-assisted methods for identifying molecules which will bind to the EPO receptor and act as an erythropoietin (EPO) mimetic. Preferred EPO mimetics identified using the method of the invention act as agonists of the EPO receptor in one or more in vitro or in vivo biological assays of EPO activity.
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  • Mambalgins, snake peptides against inflammatory and neuropathic pain through inhibition of ASIC channels

    Anne Baron   Sylvie Diochot   Miguel Salinas   Abdelkrim Alloui   Dominique Douguet   Gilles Mourier   Pascal Kessler   Enrico A. Stura   Thomas Besson   Valérie Friend   Denis Servent   Alain Eschalier   Eric Lingueglia  

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  • A simple modification of the Q-plate for parallel screening and combinatorial crystallization

    Enrico A. Stura  

    In many cases it can be advantageous to screen crystallization conditions in parallel. Tasks, such as the setting up a standard screen for several proteins simultaneously or analyzing the effect of drop size and different precipitant-protein drop ratios can be done more efficiently when drops are set up in parallel using a common reservoir. However, a real need for parallel screening becomes apparent when analyzing a large number of different complexes of the same protein, or when using methods such as stoichiometry variation screening and other combinatorial crystallization methods. Because of its large size, the Q-plate is easily adapted to the screening of a variable number of sitting drops in parallel. The drops are set up on a single glass slide placed over the same reservoir solution. The modification is simple and does not require a robot although it is well suited for automation. A record of each drop in multi-drop setup is kept as an HTML-file which allows for rapid navigation between experiments. Flexibility in the number drops makes the method effective for most combinatorial and non-combinatorial applications
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  • Crystallization of Antibodies and Antibody-Antigen Complexes

    Enrico A. Stura   Gail G. Fieser   Ian A. Wilson  

    Abstract Although many antibodies have been crystallized, the number of structures determined in both their complexed and unliganded forms remains relatively small. With the recent improvements in the use of molecular replacement (MR), the structure determination of Fabs and Fab-complex structures can proceed more rapidly, but crystallization often remains a major obstacle. Substantial improvements in methodologies have helped with the success rate in the crystallizations of Fabs and Fab-antigen complexes that are beyond previous expectations. Crystallization and structure determination have been directed mainly toward Fab fragments. The reason for this choice remains linked both to the ease with which the structure of Fabs can be determined and to the difficulties that have been presented by the crystallization of whole immunoglobulins. Such difficulty is currently believed to be due to flexibility or conformational heterogeneity of the IgG as well as the added heterogeneity from the glycosylation of the Fc fragment. Fabs share some of the same problems mainly because of the degree of heterogeneity that is the result of the proteolytic cleavage used to fragment the immunoglobulins, of the flexibility in elbow regions, and in some cases of glycosylatlon. A systematic approach to the cleavage, purification, and analysis of the resultant product can yield inmunoglobulin fragments amenable to crystallization. A rational screening of crystallization conditions with extensive use of seeding can In most cases enable progress from small microcrystalline aggregates to large X-ray-quality crystals. Such methodologies have become so effective that Fabs are now being used as tools to aid in the crystallization of other molecules that have been found difficult to crystallize by themselves.
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  • Strategies in the crystallization of glycoproteins and protein complexes

    Enrico A. Stura   Glen R. Nemerow   Ian A. Wilson  

    Modern biochemical and molecular biological techniques have provided new opportunities to investigate the structure of more complex biomolecules and have opened new paths for the crystallization of complexes. Desialation, deglycosylation and modification of glycoproteins are techniques being investigated as a means of making glycosylated proteins more amenable for crystallization. A simple solubility screen based on a limited set of precipitants has been extensively used in the comparison of various protein preparations and in the crystallization of macromolecular complexes. Antibodies, or their Fabs or Fab' fragments, can also be utilized in the crystallization of glycoproteins or other proteins which have proved difficult to crystallize by themselves. Fab complexes can provide different surfaces for lattices to form and may increase the likelihood of crystallizing a given protein. This method can be extended by the addition of an epitope tag, such as a short peptide sequence, to a protein by genetic engineering methods
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  • Applications of the streak seeding technique in protein crystallization

    Enrico A. Stura   Ian A. Wilson  

    A technique where seeds from a crystal are streaked into a protein-precipitant drop along a straight line using a rabbit whisker has been developed to determine suitable conditions for either micro- or macro-seeding. This seeding technique has also been successful in obtaining crystals using crystalline precipitate as the source of seeds, in cross-seeding from Fab-peptide complex crystals to obtain complexes with peptides of longer length, and similarly in cross-seeding from crystals of one monoclonal Fab-peptide complex to obtain crystals of a different monoclonal Fab complexed to the same peptide. This relatively simple technique should be of general applicability in macromolecular crystallizations and offers the potential to test whether new complex crystals may be obtained by seeding with the native or other complexes, to determine whether cross-seeding may be used with seeds from a related protein or to analyze the crystallization potential of different protein preparations
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  • Crystallization studies of glycosylated and unglycosylated human recombinant interleukin-2

    Enrico A. Stura   Ping Chen   Carrie M. Wilmot   Jairo H. Arevalo   Ian A. Wilson  

    Glycosylated interleukin-2 (glyIL-2) has been crystallized in two crystal forms, and unglycosylated interleukin-2 (uIL-2) has been crystallized in three forms. The glycosylated form of the human recombinant IL-2 has been crystallized from 1.9 M ammonium sulfate, pH 6.5 to 7.0 in the hexagonal space group P6222 or its enantiomorph. The crystals diffract to 2.8 .ANG. and contain two or three molecules per asymmetric unit. A second crystal form grows from 1.4 to 1.5 M ammonium sulfate on 0.2 M ammonium acetate pH 5.0-5.5, as polycrystalline rosettes which are not suitable for even a preliminary crystallographic analysis. The uIL-2 crystallizes from 1.0 to 1.7 M ammonium sulfate, 0.2 M ammonium acetate, pH 4.5-5.6 in the monoclinic space group P21, and less frequently in the orthorhombic space group P212121 from 2.5 M ammonium sulfate, pH 4.5 to 5.7. Cross-seeding uIL-2 with seeds from hexagonal crystals of glyIL-2 promotes nucleation of trigonal crystals of unglycosylated IL-2. These trigonal crystals belong to the space group P3121 or its enantiomorph, with similar cell dimensions to the glyIL-2 hexagonal crystals.
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  • Crystallization of murine major histocompatibility complex class I H-2Kb with single peptides

    Enrico A. Stura   Masazumi Matsumura   Daved H. Fremont   Yutaka Saito   Per A. Peterson   Lan A. Wilson  

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  • Crystal structure and functional domains of the Mambaquaretin-1, a vasopressin type 2 receptor peptide inhibitor for kidney cysts treatment

    Laura Droctové   Justyna Ciolek   Manon Lancien   Enrico A. Stura   Laura Vera   Nicolas Floquet   Ralph Witzgall   Bernard Mouillac   Christiane Mendre   Gilles Mourier   Denis Servent   Nicolas Gilles  

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  • A Tribute to Enrico Drioli

    Giorno, Lidietta   Paul, Donald R.  

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  • A Tribute to Enrico Drioli

    Giorno, Lidietta; Paul, Donald R.  

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  • Potere e limiti della conoscenza da Enrico di Gand a Enrico di Harclay

    Arezzo   Anna  

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  • Potere e limiti della conoscenza da Enrico di Gand a Enrico di Harclay

    Arezzo, Anna  

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  • A Review of Enrico Moretti's The New Geography of Jobs

    Glaeser, Edward  

    Why is prosperity distributed so unevenly across America's metropolitan areas? While population growth has gone disproportionately towards the Sunbelt, high-skill areas have experienced the strongest income growth since 1970. Gaps between more and less educated areas were modest forty years ago, but they have become quite large, and far larger than would be predicted solely by the general rise in the returns to skill. Unemployment rates during the recent recession were also strongly correlated with area level education. This essay reviews Enrico Moretti's The New Geography of Jobs, which both describes and explains these significant regional trends.
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