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  1. Article ; Online: Generation of a Soluble Form of Human Endoglin Fused to Green Fluorescent Protein

    Lidia Ruiz-Llorente / M. Cristina Vega / Francisco J. Fernández / Carmen Langa / Nicholas W. Morrell / Paul D. Upton / Carmelo Bernabeu

    International Journal of Molecular Sciences, Vol 22, Iss 11282, p

    2021  Volume 11282

    Abstract: Endoglin (Eng, CD105) is a type I membrane glycoprotein that functions in endothelial cells as an auxiliary receptor for transforming growth factor ... ...

    Abstract Endoglin (Eng, CD105) is a type I membrane glycoprotein that functions in endothelial cells as an auxiliary receptor for transforming growth factor <math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">β</mi></semantics></math> (TGF- <math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">β</mi></semantics></math> )/bone morphogenetic protein (BMP) family members and as an integrin ligand, modulating the vascular pathophysiology. Besides the membrane-bound endoglin, there is a soluble form of endoglin (sEng) that can be generated by the action of the matrix metalloproteinase (MMP)-14 or -12 on the juxtamembrane region of its ectodomain. High levels of sEng have been reported in patients with preeclampsia, hypercholesterolemia, atherosclerosis and cancer. In addition, sEng is a marker of cardiovascular damage in patients with hypertension and diabetes, plays a pathogenic role in preeclampsia, and inhibits angiogenesis and tumor proliferation, migration, and invasion in cancer. However, the mechanisms of action of sEng have not yet been elucidated, and new tools and experimental approaches are necessary to advance in this field. To this end, we aimed to obtain a fluorescent form of sEng as a new tool for biological imaging. Thus, we cloned the extracellular domain of endoglin in the pEGFP-N1 plasmid to generate a fusion protein with green fluorescent protein (GFP), giving rise to pEGFP-N1/Eng.EC. The recombinant fusion protein was characterized by transient and stable transfections in CHO-K1 cells using fluorescence microscopy, SDS-PAGE, immunodetection, and ELISA techniques. Upon transfection with pEGFP-N1/Eng.EC, fluorescence was readily detected in cells, indicating that the GFP contained in the recombinant protein was properly folded into the cytosol. Furthermore, as evidenced by Western blot analysis, the secreted fusion protein yielded ...
    Keywords endoglin ; GFP ; soluble endoglin ; fusion protein ; recombinant protein ; fluorescence ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Subject code 500
    Language English
    Publishing date 2021-10-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: The crystal structure of iC3b-CR3 αI reveals a modular recognition of the main opsonin iC3b by the CR3 integrin receptor

    Francisco J. Fernández / Jorge Santos-López / Rubén Martínez-Barricarte / Javier Querol-García / Héctor Martín-Merinero / Sergio Navas-Yuste / Martin Savko / William E. Shepard / Santiago Rodríguez de Córdoba / M. Cristina Vega

    Nature Communications, Vol 13, Iss 1, Pp 1-

    2022  Volume 16

    Abstract: Complement activation on foreign cell surfaces leads to the generation of complement opsonins, which activate complement receptor type 3 (CR3) and pathogen clearance by macrophages. Here, the authors reveal structural basis of the interaction between ... ...

    Abstract Complement activation on foreign cell surfaces leads to the generation of complement opsonins, which activate complement receptor type 3 (CR3) and pathogen clearance by macrophages. Here, the authors reveal structural basis of the interaction between human opsonin iC3b and the von Willebrand A inserted domain of the α chain of CR3.
    Keywords Science ; Q
    Language English
    Publishing date 2022-04-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Híbridos dobles de maíz formados con líneas mejoradas por selección gamética y retrocruza.

    Enrique Navarro / Gustavo Burciaga / Salvador González / M. Cristina Vega / Regino Morones / Elias Sandoval

    Agronomía Mesoamericana, Vol 8, Iss

    2016  Volume 2

    Abstract: El presente trabajo involucra la evaluación de 1040 híbridos dobles formados a partir de cruzas simples mejoradas a través del método de selección gamética y retrocruza, las mismas que fueron cruzadas con cinco probadores y asignadas a ocho experimentos, ...

    Abstract El presente trabajo involucra la evaluación de 1040 híbridos dobles formados a partir de cruzas simples mejoradas a través del método de selección gamética y retrocruza, las mismas que fueron cruzadas con cinco probadores y asignadas a ocho experimentos, de acuerdo a la naturaleza de las cruzas simples. La evaluación se llevó a cabo en 1991 en Gómez Palacio, Durango, México, con los siguientes objetivos: 1) Evaluar el comportamiento de los híbridos dobles y 2) Determinar los caracteres agrónomicos de mayor importancia en la variación observada. Para rendimiento de grano, sobresalieron por su productividad los híbridos dobles formados con cruzas simples mejoradas por selección gamética, con rendimientos de hasta 11,6 t/ha, superando en un 23% al mejor del procedimiento de retrocruza, lo anterior es un reflejo del reducido por ciento de mazorcas podridas observado en selección gamética. Las cruzas simples mejoradas por retrocruzamiento fueron más precoces, de menor altura y con menos problemas de acame de raíz y tallo. Los mejores probadores fueron el 258-18-19 x MLS4-1 y el 232 x 255-18-19, ya que al combinarse con algunas cruzas simples sus rendimientos de grano excedieron las 14 t/ha. Los máximos rendimientos de grano estuvieron asociados con valores altos de ACG en selección gamética. El análisis multivariado detectó que más de las dos terceras partes de la variación de los híbridos dobles es explicada por rendimiento de grano limpio, altura de mazorca y floración masculina- floración femenina.
    Keywords Agriculture ; S
    Language Spanish
    Publishing date 2016-06-01T00:00:00Z
    Publisher Universidad de Costa Rica
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Structures of T7 bacteriophage portal and tail suggest a viral DNA retention and ejection mechanism

    Ana Cuervo / Montserrat Fàbrega-Ferrer / Cristina Machón / José Javier Conesa / Francisco J. Fernández / Rosa Pérez-Luque / Mar Pérez-Ruiz / Joan Pous / M. Cristina Vega / José L. Carrascosa / Miquel Coll

    Nature Communications, Vol 10, Iss 1, Pp 1-

    2019  Volume 11

    Abstract: The genome of double-stranded DNA bacteriophages is delivered to host cells through the viral portal and tail structures. Here the authors describe structures of the bacteriophage T7 portal and the 1.5 MDa tail complex formed by the portal protein, ... ...

    Abstract The genome of double-stranded DNA bacteriophages is delivered to host cells through the viral portal and tail structures. Here the authors describe structures of the bacteriophage T7 portal and the 1.5 MDa tail complex formed by the portal protein, adaptor protein and nozzle, providing insight into how the portal and tail machinery work during DNA packaging and ejection.
    Keywords Science ; Q
    Language English
    Publishing date 2019-08-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Structures of T7 bacteriophage portal and tail suggest a viral DNA retention and ejection mechanism

    Ana Cuervo / Montserrat Fàbrega-Ferrer / Cristina Machón / José Javier Conesa / Francisco J. Fernández / Rosa Pérez-Luque / Mar Pérez-Ruiz / Joan Pous / M. Cristina Vega / José L. Carrascosa / Miquel Coll

    Nature Communications, Vol 10, Iss 1, Pp 1-

    2019  Volume 11

    Abstract: The genome of double-stranded DNA bacteriophages is delivered to host cells through the viral portal and tail structures. Here the authors describe structures of the bacteriophage T7 portal and the 1.5 MDa tail complex formed by the portal protein, ... ...

    Abstract The genome of double-stranded DNA bacteriophages is delivered to host cells through the viral portal and tail structures. Here the authors describe structures of the bacteriophage T7 portal and the 1.5 MDa tail complex formed by the portal protein, adaptor protein and nozzle, providing insight into how the portal and tail machinery work during DNA packaging and ejection.
    Keywords Science ; Q
    Language English
    Publishing date 2019-08-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Mechanical network in titin immunoglobulin from force distribution analysis.

    Wolfram Stacklies / M Cristina Vega / Matthias Wilmanns / Frauke Gräter

    PLoS Computational Biology, Vol 5, Iss 3, p e

    2009  Volume 1000306

    Abstract: The role of mechanical force in cellular processes is increasingly revealed by single molecule experiments and simulations of force-induced transitions in proteins. How the applied force propagates within proteins determines their mechanical behavior yet ...

    Abstract The role of mechanical force in cellular processes is increasingly revealed by single molecule experiments and simulations of force-induced transitions in proteins. How the applied force propagates within proteins determines their mechanical behavior yet remains largely unknown. We present a new method based on molecular dynamics simulations to disclose the distribution of strain in protein structures, here for the newly determined high-resolution crystal structure of I27, a titin immunoglobulin (IG) domain. We obtain a sparse, spatially connected, and highly anisotropic mechanical network. This allows us to detect load-bearing motifs composed of interstrand hydrogen bonds and hydrophobic core interactions, including parts distal to the site to which force was applied. The role of the force distribution pattern for mechanical stability is tested by in silico unfolding of I27 mutants. We then compare the observed force pattern to the sparse network of coevolved residues found in this family. We find a remarkable overlap, suggesting the force distribution to reflect constraints for the evolutionary design of mechanical resistance in the IG family. The force distribution analysis provides a molecular interpretation of coevolution and opens the road to the study of the mechanism of signal propagation in proteins in general.
    Keywords Biology (General) ; QH301-705.5
    Subject code 612
    Language English
    Publishing date 2009-03-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: Insights into the inhibited form of the redox-sensitive SufE-like sulfur acceptor CsdE.

    Esther Peña-Soler / Juan Aranda / Miguel López-Estepa / Sara Gómez / Fernando Garces / Miquel Coll / Francisco J Fernández / Iñaki Tuñon / M Cristina Vega

    PLoS ONE, Vol 12, Iss 10, p e

    2017  Volume 0186286

    Abstract: Sulfur trafficking in living organisms relies on transpersulfuration reactions consisting in the enzyme-catalyzed transfer of S atoms via activated persulfidic S across protein-protein interfaces. The recent elucidation of the mechanistic basis for ... ...

    Abstract Sulfur trafficking in living organisms relies on transpersulfuration reactions consisting in the enzyme-catalyzed transfer of S atoms via activated persulfidic S across protein-protein interfaces. The recent elucidation of the mechanistic basis for transpersulfuration in the CsdA-CsdE model system has paved the way for a better understanding of its role under oxidative stress. Herein we present the crystal structure of the oxidized, inactivated CsdE dimer at 2.4 Å resolution. The structure sheds light into the activation of the Cys61 nucleophile on its way from a solvent-secluded position in free CsdE to a fully extended conformation in the persulfurated CsdA-CsdE complex. Molecular dynamics simulations of available CsdE structures allow to delineate the sequence of conformational changes underwent by CsdE and to pinpoint the key role played by the deprotonation of the Cys61 thiol. The low-energy subunit orientation in the disulfide-bridged CsdE dimer demonstrates the likely physiologic relevance of this oxidative dead-end form of CsdE, suggesting that CsdE could act as a redox sensor in vivo.
    Keywords Medicine ; R ; Science ; Q
    Subject code 500
    Language English
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article: Structural and functional characterization of a highly stable endo-β-1,4-xylanase from Fusarium oxysporum and its development as an efficient immobilized biocatalyst

    Gómez, Sara / Asia M. Payne / Francisco J. Fernandez / Gavin C. Fox / M. Cristina Vega / Martin Savko / William E. Shepard

    Biotechnology for biofuels. 2016 Dec., v. 9, no. 1

    2016  

    Abstract: BACKGROUND: Replacing fossil fuel with renewable sources such as lignocellulosic biomass is currently a promising alternative for obtaining biofuel and for fighting against the consequences of climate change. However, the recalcitrant structure of ... ...

    Abstract BACKGROUND: Replacing fossil fuel with renewable sources such as lignocellulosic biomass is currently a promising alternative for obtaining biofuel and for fighting against the consequences of climate change. However, the recalcitrant structure of lignocellulosic biomass residues constitutes a major limitation for its widespread use in industry. The efficient hydrolysis of lignocellulosic materials requires the complementary action of multiple enzymes including xylanases and β-xylosidases, which are responsible for cleaving exo- and endoxylan linkages, that release oligocarbohydrates that can be further processed by other enzymes. RESULTS: We have identified the endo-β-1,4-xylanase Xyl2 from Fusarium oxysporum as a promising glycoside hydrolase family 11 enzyme for the industrial degradation of xylan. To characterize Xyl2, we have cloned the synthetic optimized gene and expressed and purified recombinant Xyl2 to homogeneity, finally obtaining 10 mg pure Xyl2 per liter of culture. The crystal structure of Xyl2 at 1.56 Å resolution and the structure of a methyl-xylopyranoside Xyl2 complex at 2.84 Å resolution cast a highly detailed view of the active site of the enzyme, revealing the molecular basis for the high catalytic efficiency of Xyl2. The kinetic analysis of Xyl2 demonstrates high xylanase activity and non-negligible β-xylosidase activity under a variety of experimental conditions including alkaline pH and elevated temperature. Immobilizing Xyl2 on a variety of solid supports enhances the enzymatic properties that render Xyl2 a promising industrial biocatalyst, which, together with the detailed structural data, may establish Xyl2 as a platform for future developments of industrially relevant xylanases. CONCLUSIONS: F. oxysporum Xyl2 is a GH11 xylanase which is highly active in free form and immobilized onto a variety of solid supports in a wide pH range. Furthermore, immobilization of Xyl2 on certain supports significantly increases its thermal stability. A mechanistic rationale for Xyl2's remarkable catalytic efficiency at alkaline pH is proposed on the basis of two crystallographic structures. Together, these properties render Xyl2 an attractive biocatalyst for the sustainable industrial degradation of xylan.
    Keywords active sites ; biocatalysts ; biofuels ; biomass ; catalytic activity ; climate change ; crystal structure ; fossil fuels ; Fusarium oxysporum ; gene expression ; glycosides ; hydrolysis ; industry ; kinetics ; lignocellulose ; pH ; temperature ; thermal stability ; xylan ; xylan 1,4-beta-xylosidase
    Language English
    Dates of publication 2016-12
    Size p. 191.
    Publishing place BioMed Central
    Document type Article
    ZDB-ID 2421351-2
    ISSN 1754-6834
    ISSN 1754-6834
    DOI 10.1186/s13068-016-0605-z
    Database NAL-Catalogue (AGRICOLA)

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  9. Article ; Online: Crystal structure, SAXS and kinetic mechanism of hyperthermophilic ADP-dependent glucokinase from Thermococcus litoralis reveal a conserved mechanism for catalysis.

    Jaime Andrés Rivas-Pardo / Alejandra Herrera-Morande / Victor Castro-Fernandez / Francisco J Fernandez / M Cristina Vega / Victoria Guixé

    PLoS ONE, Vol 8, Iss 6, p e

    2013  Volume 66687

    Abstract: ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions ... ...

    Abstract ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions associated with ligand binding and whether they are meaningful to the function of the enzyme. We used the ADP-dependent glucokinase from Thermococcus litoralis as a model to investigate the conformational changes observed in X-ray crystallographic structures upon substrate binding and to compare them with those determined in solution in order to understand their interplay with the glucokinase function. Initial velocity studies indicate that catalysis follows a sequential ordered mechanism that correlates with the structural transitions experienced by the enzyme in solution and in the crystal state. The combined data allowed us to resolve the open-closed conformational transition that accounts for the complete reaction cycle and to identify the corresponding clusters of aminoacids residues responsible for it. These results provide molecular bases for a general mechanism conserved across the ADP-dependent kinase family.
    Keywords Medicine ; R ; Science ; Q
    Subject code 540
    Language English
    Publishing date 2013-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: Correction

    Miguel López-Estepa / Ana Ardá / Martin Savko / Adam Round / William E Shepard / Marta Bruix / Miquel Coll / Francisco J Fernández / Jesús Jiménez-Barbero / M Cristina Vega

    PLoS ONE, Vol 10, Iss 7, p e

    The Crystal Structure and Small-Angle X-Ray Analysis of CsdL/TcdA Reveal a New tRNA Binding Motif in the MoeB/E1 Superfamily.

    2015  Volume 0134070

    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2015-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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