<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>0379-3982</journal-id>
<journal-title><![CDATA[Revista Tecnología en Marcha]]></journal-title>
<abbrev-journal-title><![CDATA[Tecnología en Marcha]]></abbrev-journal-title>
<issn>0379-3982</issn>
<publisher>
<publisher-name><![CDATA[Instituto Tecnológico de Costa Rica]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0379-39822016000400078</article-id>
<article-id pub-id-type="doi">10.18845/tm.v29i4.3039</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Producción heteróloga y caracterización bioquímica del procoagulante humano Factor VIII para ensayos de cristalización de macromoléculas proteicas]]></article-title>
<article-title xml:lang="en"><![CDATA[Heterologous production and biochemical characterization of human pro-coagulation Factor VIII for crystallization screening of protein macromolecules]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Hernández-Carvajal]]></surname>
<given-names><![CDATA[Erick]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Arce-Solano]]></surname>
<given-names><![CDATA[Silvia]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Mena-Aguilar]]></surname>
<given-names><![CDATA[Didier]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Fuentes-Prior]]></surname>
<given-names><![CDATA[Pablo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Hospital de la Santa Creu i Sant Pau Institut d&#8217;Investigació Biomèdica ]]></institution>
<addr-line><![CDATA[Barcelona ]]></addr-line>
<country>España</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>29</volume>
<numero>4</numero>
<fpage>78</fpage>
<lpage>91</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_arttext&amp;pid=S0379-39822016000400078&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_abstract&amp;pid=S0379-39822016000400078&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_pdf&amp;pid=S0379-39822016000400078&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen A pesar de la relevancia médica y el impacto económico en los sistemas de salud, las enfermedades cardiovasculares, tales como infartos cardíacos y accidentes cerebrovasculares, siguen siendo la principal causa de morbilidad y mortalidad en los países en vías de desarrollo. Esto se debe a que las bases estructurales y funcionales de los procesos de formación de los coágulos sanguíneos o trombos solo se conocen de forma incompleta. La trombina juega un papel esencial en estos procesos y los fundamentos atómico-moleculares de su interacción con otros factores que participan en el proceso de coagulación son poco conocidos, en particular el reconocimiento de importantes sustratos como los factores V y VIII, así como el receptor de plaquetas PAR1. Dada la importancia de estos factores, en esta investigación se produjeron fragmentos del factor VIII humano (FVIII) y se caracterizaron bioquímicamente para realizar ensayos de cristalización de complejos FVIII·Trombina. Para ello, (1) se sobreexpresaron heterólogamente los conectores ácidos entre los dominios del factor VIII (denominados FVIIIa1, FVIIIa2 y FVIIIa3), (2) se purificaron y caracterizaron estos fragmentos recombinantes, (3) se formaron sus complejos con la trombina y (4) se inició la búsqueda de las condiciones de cristalización de estos complejos proteicos. La producción del FVIII, y en particular la determinación de las condiciones en las que crecen cristales del tamaño y calidad apropiados, son un auténtico cuello de botella en los estudios de estructura-función, por ello se considera que la optimización de estos procesos permitirá obtener un mayor número de cristales de proteína de calidad adecuada para futuros estudios de difracción de rayos X.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract In spite of their medical relevance and economic impact on public health service systems, cardiovascular diseases, among them myocardial and cerebral infarctions, remain the main cause of morbidity and mortality in developed and developing countries. One of the main reasons for this situation is the incomplete structural and functional knowledge of the processes that ultimately lead to blood clot formation. Thrombin plays a key role in these processes, but its molecular interactions with other clotting factors are still only poorly understood, in particular those with critically important substrates such as factor VIII and the thrombin receptor on platelets, PAR1. Given the importance of these clotting factors, we have produced and characterized factor VIII fragments (FVIII) to search for crystallization conditions of FVIII·Thrombin complexes. To achieve our aims, we (1) overexpressed the acidic inter-domain linkers of human factor VIII (FVIIIa1, FVIIIa2 and FVIIIa3), as heterologous proteins; (2) purified and characterized these recombinant forms, (3) formed the binary protein complexes between thrombin and these recombinant fragments, and (4) started crystallization trials of these complexes. The FVIII production, and in particular, the identification of conditions in which crystals of appropriate size and quality for structure determination grow, are a &#8220;bottleneck&#8221; in structure-and-function projects. Therefore, the optimization of these processes will allow us to obtain well-ordered protein crystals for X-ray diffraction studies.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Enfermedad cardiovascular]]></kwd>
<kwd lng="es"><![CDATA[trombina]]></kwd>
<kwd lng="es"><![CDATA[FVIII]]></kwd>
<kwd lng="es"><![CDATA[cristalización de proteínas]]></kwd>
<kwd lng="es"><![CDATA[difracción de rayos X]]></kwd>
<kwd lng="en"><![CDATA[Cardiovascular disease]]></kwd>
<kwd lng="en"><![CDATA[thrombin]]></kwd>
<kwd lng="en"><![CDATA[FVIII]]></kwd>
<kwd lng="en"><![CDATA[protein crystallization]]></kwd>
<kwd lng="en"><![CDATA[X-ray diffraction]]></kwd>
</kwd-group>
</article-meta>
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<surname><![CDATA[Sheehan]]></surname>
<given-names><![CDATA[J.P]]></given-names>
</name>
</person-group>
<article-title xml:lang=""><![CDATA[The factor IXa heparin-binding exosite is a cofactor interactive site: mechanism for antithrombin-independent inhibition of intrinsic tenase by heparin]]></article-title>
<source><![CDATA[Biochemistry]]></source>
<year>2005</year>
<volume>44</volume>
<page-range>3615-25</page-range></nlm-citation>
</ref>
</ref-list>
</back>
</article>
